Objective: Our objective was to determine growth rates, body composition, and pubertal development of replacement gilts fed diets with different ratios of standardized ileal digestible (SID) lysine to ME. Materials and Methods: Diets with low, medium, and high ratios of SID lysine to ME (grower: 2.3, 2.6, and 2.8; and, finisher; 1.7, 1.9, and 2.1 g/Mcal) were fed from 100 to 200 d of age, after which gilts were moved from the gilt development unit to sow farms. Boar exposure and estrus detection began at 160 d of age and continued until first detected estrus. Estimates of BW and body composition were determined at 100, 142, 160, and 200 d of age and at puberty. Results and Discussion: Body weights and growth rates were reduced (P < 0.05) as dietary SID lysine-to-ME ratio decreased. Greater SID lysine-to-ME ratios increased the number of gilts that exhibited estrus upon boar exposure, increased the number of gilts with a spontaneous first estrus, reduced the number of gilts requiring P.G.600 (Merck Animal Health, Kenilworth, NJ), and decreased age at first estrus (P < 0.05). Slower growing gilts that weighed less with less backfat were more likely to require P.G.600 to induce puberty (P < 0.05). Implications and Applications: Reducing SID lysine-to-ME ratios in gilt diets can increase the number of gilts within optimal BW range at first estrus, but overall pubertal development is delayed if ratios are reduced below 2.8 and 2.1 g of SID lysine to megacalorie of ME in grower and finisher diets, respectively.

Maternal effects on development are profound. Together, genetic and epigenetic maternal effects define the developmental trajectory of progeny and, ultimately, offspring phenotype. Maternally provisioned environmental conditions and signals affect conceptus, fetoplacental and postnatal development from the time of conception until weaning. In the pig, reproductive tract development is completed postnatally. Porcine uterine growth and uterine endometrial development occur in an ovary-independent manner between birth (postnatal day = PND 0) and PND 60. Milk-borne bioactive factors (MbFs), exemplified by relaxin, communicated from lactating dam to nursing offspring via a lactocrine mechanism, represent an important source of extraovarian uterotrophic support in the neonatal pig. Lactocrine deficiency from birth affects both the neonatal porcine uterine developmental program and trajectory of uterine development, with lasting consequences for endometrial function and uterine capacity in adult female pigs. The potential lactocrine signaling window extends from birth until the time of weaning. However, it is likely that the maternal lactocrine programming window – that period when MbFs communicated to nursing offspring have the greatest potential to affect critical organizational events in the neonate – encompasses a comparatively short period of time within 48 h of birth. Lactocrine deficiency from birth was associated with altered patterns of endometrial gene expression in neonatally lactocrine-deficient adult gilts during a critical period for conceptus–endometrial interaction on pregnancy day 13, and with reduced litter size, estimated at 1.4 pigs per litter, with no effect of parity. Data were interpreted to indicate that reproductive performance of female pigs that do not receive sufficient colostrum from birth is permanently impaired. Observations to date suggest that lactocrine-dependent maternal effects program postnatal development of the porcine uterus, endometrial functionality and uterine capacity. In this context, reproductive management strategies and husbandry guidelines should be refined to ensure that such practices promote environmental conditions that will optimize uterine capacity and fecundity. This will entail careful consideration of factors affecting lactation, the quality and abundance of colostrum/milk, and practices that will afford neonatal pigs with the opportunity to nurse and consume adequate amounts of colostrum.

Maternal effects on development can program cell fate and dictate offspring phenotype. Such effects do not end at birth, but extend into postnatal life through signals communicated from mother to offspring in first milk (colostrum). Transmission of bioactive factors from mother to offspring as a specific consequence of nursing defines a lactocrine mechanism. The female reproductive tract is not fully formed at birth (postnatal day = PND 0). Data for ungulates and mice indicate that disruption of development during neonatal life can have lasting effects on the form and function of uterine tissues. Uterine growth and histogenesis proceed in an ovary-independent manner shortly after birth, suggesting that extra-ovarian inputs are important in this process. Data for the pig indicate that lactocrine signals communicated within 12 to 48 h from birth constitute one source of such uterotrophic support. Disruption of lactocrine signaling, either naturally, by limited colostrum consumption, or experimentally, by milk replacer feeding, alters neonatal porcine uterine development and can have negative consequences for reproductive performance in adults. Substantial differences in endometrial and uterine gene expression between colostrum- and replacer-fed gilts were evident by PND 2, when RNA sequencing revealed over 800 differentially expressed, lactocrine-sensitive genes. Lactocrine-sensitive biological processes identified through transcriptomic studies and integrated microRNA-mRNA pathway analyses included those associated with both cell-cell and ESR1 signaling, and tissue development. Evidence for the pig indicates that colostrum consumption and lactocrine signaling are required to establish a normal uterine developmental program and optimal uterine developmental trajectory.

Nursing for 2 d from birth supports neonatal porcine uterine and cervical development. However, it is not clear how timing or duration of lactocrine signaling from birth (postnatal day = PND 0) affects development of neonatal female reproductive tract tissues. Therefore, studies were conducted to determine effects of age at first nursing and duration of nursing from birth on specific elements of the matrix metalloproteinase (MMP)/tissue inhibitor of metalloproteinase (TIMP) system in uterine and cervical tissues at PND 2. When nursing was initiated at 0 h or 30 min of age, targeted proteins, including proMMP9 and MMP9, were detected in uterine and cervical tissues on PND 2, as was uterine TIMP1. However, these proteins were undetectable when nursing was delayed for 12 h and when gilts were fed milk replacer for 48 h from birth. Increasing the duration of nursing from 30 min to 12 h from birth increased uterine (P < 0.05) and cervical (P < 0.001) MMP9 levels to those observed in gilts nursed for 48 h. Similarly, uterine TIMP1 levels increased with duration of nursing. Uterine MMP2 levels were detectable but unaffected by age at first nursing or duration of nursing from birth. Uterine MMP2 and MMP9 activities, monitored by zymography, reflected immunoblotting data. Results provide evidence for the utility of MMP9 and TIMP1 as markers of age- and lactocrine-sensitive porcine female reproductive tract development.

The idea, suggested by Charles Stockard in 1921, that there are critical periods during which disruption of the developmental program can have lasting effects on the form and function of tissues, organs and even organisms, is now over 90 yr old. Twenty years later, Conrad Waddington coined the term ‘epigenotype’ to describe mechanisms regulating development, and ultimately proposed that cell fate could be determined or programmed by changes in the ‘epigenetic landscape’. The ‘thrifty phenotype hypothesis’, proposed by David Barker in 1990, advanced these ideas, suggesting that development includes programming by the environment in fetal and infant life, and that disruption of development during critical organizational periods can have life-long consequences. Much attention has been focused on identification of critical organizational periods during fetal life. However, it is important to remember that developmental plasticity and, consequently, susceptibility of tissues to developmental disruption does not end at birth but can extend into the postnatal period. Data for laboratory as well as large domestic animals indicate that targeted disruption of steroid hormone-sensitive developmental events during early neonatal life can alter the form and/or function of adult reproductive tract tissues, including the uterus. The uterine gland knockout (UGKO) phenotype, characterized by the absence of endometrial glands in adult sheep, cattle and mice exposed neonatally to steroidal compounds, provides the prototypical example of such programming effects. Factors affecting reproductive development can also be communicated from mother to offspring in first milk (colostrum) via a lactocrine mechanism. In the pig, imposition of a lactocrine-null state by substituting milk replacer for colostrum for 2 d from birth altered patterns of uterine gene expression and retarded endometrial gland development. Transcriptomic analysis of neonatal uterine gene expression in nursed as compared to replacer-fed gilts on postnatal Day 2 revealed over 800 differentially expressed, lactocrine-sensitive genes. A subset of non-coding microRNAs targeting over 100 lactocrine-sensitive uterine transcripts was recently identified, indicating potential for further epigenetic regulation of uterine development. Evidence that minimal colostrum consumption at birth is associated with reduced lifetime fecundity in adult sows supports the idea that lactocrine programming is a factor affecting reproductive efficiency. Data for the mouse indicating lactocrine effects on hippocampal development and memory, and for primates indicating lactocrine effects on offspring temperament and growth support a role for lactocrine regulation of postnatal development. Potential for transgenerational epigenetic effects emphasizes the importance of understanding the biology of postnatal programming and reproductive development.

Maternal contributions to development begin at conception. Prenatal conditions that evolve in utero through the course of gestation define the environment in which embryogenesis and fetoplacental development occur. Genotype notwithstanding, maternal effects on development from the time of conception can program cell fate and dictate offspring phenotype as defined by various aspects of performance and health, including fertility and fecundity. Maternal effects on development do not end at birth, but extend into postnatal life through signals communicated from mother to offspring in first milk (colostrum). Transmission of bioactive factors from mother to offspring as a specific consequence of nursing defines a lactocrine mechanism. The female reproductive tract (FRT) is not fully formed at birth. Data for both ungulate species and mice indicate that disruption of the developmental program during critical organizational periods of neonatal life can have lasting effects on the form and function of FRT tissues, including the uterus. Radial patterning of the uterine wall, reflected by differentiation and proliferation of nascent endometrial glands, is a postnatal event in most mammals. Both uterine growth and histogenesis proceed in an ovary-independent manner shortly after birth, suggesting that extra-ovarian inputs are important in this process. Data for the pig indicate that lactocrine signals constitute one source of such uterotrophic support. Disruption of lactocrine signaling by feeding gilts porcine milk replacer instead of colostrum for 2 d from birth (postnatal day = PND 0) retarded uterine gland genesis by PND 14. Differences in endometrial and whole uterine gene expression patterns between colostrum- and replacer-fed gilts were evident by PND 2, when RNA sequencing revealed over 800 differentially expressed, lactocrine-sensitive genes. Organizationally relevant, lactocrine-sensitive processes, pathways, and networks identified through transcriptomic studies included cell adhesion, cell-cell signaling, cytokine-receptor interactions, integrin cell surface interactions, ESR1 and Hedgehog signaling, and the plasminogen activating network. Lactocrine-sensitive expression of nine microRNAs with 115 potential mRNA targets was also identified. Results provide evidence of lactocrine-mediated, epigenetic effects on multiple elements of the uterine developmental program. A single oral dose of colostrum given at birth affects endometrial cell behaviors associated with uterine wall development by 12 h postnatal. Evidence that minimal colostrum consumption at birth is associated with reduced lifetime fecundity in adult sows indicates that lactocrine programming can affect reproductive efficiency. Data support a role for lactocrine signaling in regulation of postnatal reproductive tract development and function.

Prepubertal exposure of the developing ovaries and reproductive tract (RT) to estrogen or xenoestrogens can have acute and long-term consequences that compromise the reproductive performance of cattle. This research examined effects of the selective estrogen receptor modulator tamoxifen (TAM) on gene and protein abundance in prepubertal ovaries and RT, with a particular focus on signaling pathways that affect morphology. Tamoxifen was administered to Holstein heifer calves (n=8) daily (0.3mg/kg subcutaneously) from 28 to 120 d of age, when tissues were collected. Control calves (n=7) received an equal volume of excipient. Weight, gross measurements, and samples of reproductive tissues were collected, and protein and mRNA were extracted from snap-frozen samples of vagina, cervix, uterus, oviduct, ovary, and liver. Neither estradiol nor insulin-like growth factor I (IGFI) concentrations in the serum were affected by TAM treatment. Tamoxifen treatment reduced ovarian weight independently from effects on antral follicle populations, as there was no difference in visible antral follicle numbers on the day of collection. Estrogen receptor α (ESR1) and β (ESR2) mRNA, ESR1 protein, IGFI, progesterone receptor, total growth hormone receptor, WNT4, WNT5A, and WNT7A mRNA, in addition to mitogen-activated protein kinase (MAPK) and phosphorylated MAPK proteins were affected differently depending on the tissue examined. However, neither IGFI receptor mRNA nor protein abundance were affected by TAM treatment. Results indicate that reproductive development in prepubertal Holstein heifer calves is TAM-sensitive, and that bovine RT and ovarian development are supported, in part, by estrogen receptor-dependent mechanisms during the period studied here. Potential long-term consequences of such developmental disruption remain to be defined.

Lactocrine signaling is defi ned as transmission of bioactive factors from mother to offspring as a consequence of nursing. Lactocrine transmission of signaling molecules may be an evolutionarily conserved process through which bioactive factors necessary for support of neonatal development are delivered postnatally. Dependence on maternal resources for development in eutherian mammals extends into neonatal life for at least that period of time when nutrition is obtained solely from fi rst milk (i.e., colostrum). Data for the pig (Sus scrofa domesticus) provide evidence of lactocrine mediated effects on development of the female reproductive tract and other somatic tissues. Porcine uterine gland development, an estrogen receptor-alpha (ESR1)dependent process, begins within 2 d of birth [postnatal day (PND) 0]. A lactocrine-driven, ESR1-mediated process was proposed as a regulatory mechanism governing onset of uterine gland development and endometrial maturation in the neonatal pig. Gilts maintained in a lactocrine-null state for 2 d from birth by milk-replacer feeding displayed altered patterns of endometrial gene expression and retarded uterine gland development by PND 14. In lactocrine-null gilts, inhibition of endometrial and cervical ESR1 and vascular endothelial growth factor (VEGFA) expression observed on PND 2 persisted to PND 14, even after gilts were returned to nursing on PND 2. Collectively, data support a role for lactocrine signaling in regulation of critical neonatal developmental events. Maternal lactocrine programming of postnatal development may help to insure healthy developmental outcomes. A systems biology approach will be required to defi ne and understand mechanistic dynamics of lactocrine signaling events that may ultimately connect genotype to phenotype and establish the parameters of reproductive potential.

Lactocrine signaling is defined as transmission of bioactive factors from mother to offspring as a consequence of nursing. Lactocrine transmission of signaling molecules may be an evolutionarily conserved process through which bioactive factors necessary for support of neonatal development are delivered postnatally. Dependence on maternal resources for development in eutherian mammals extends into neonatal life for at least that period of time when nutrition is obtained solely from first milk (i.e., colostrum). Data for the pig (Sus scrofa domesticus) provide evidence of lactocrine mediated effects on development of the female reproductive tract and other somatic tissues. Porcine uterine gland development, an estrogen receptor-alpha (ESR1)-dependent process, begins within 2 d of birth [postnatal day (PND) 0]. A lactocrine-driven, ESR1-mediated process was proposed as a regulatory mechanism governing onset of uterine gland development and endometrial maturation in the neonatal pig. Gilts maintained in a lactocrine-null state for 2 d from birth by milk-replacer feeding displayed altered patterns of endometrial gene expression and retarded uterine gland development by PND 14. In lactocrine-null gilts, inhibition of endometrial and cervical ESR1 and vascular endothelial growth factor (VEGFA) expression observed on PND 2 persisted to PND 14, even after gilts were returned to nursing on PND 2. Collectively, data support a role for lactocrine signaling in regulation of critical neonatal developmental events. Maternal lactocrine programming of postnatal development may help to insure healthy developmental outcomes. A systems biology approach will be required to define and understand mechanistic dynamics of lactocrine signaling events that may ultimately connect genotype to phenotype and establish the parameters of reproductive potential.

Xenomitochondrial mice, harboring evolutionarily divergent Mus terricolor mitochondrial DNA (mtDNA) on a Mus musculus domesticus nuclear background (B6NTac(129S6)-mtM. terricolor/Capt; line D7), were subjected to molecular and phenotypic analyses. No overt in vivo phenotype was identified in contrast to in vitro xenomitochondrial cybrid studies. Microarray analyses revealed differentially expressed genes in xenomitochondrial mice, though none were directly involved in mitochondrial function. qRT-PCR revealed upregulation of mt-Co2 in xenomitochondrial mice. These results illustrate that cellular compensatory mechanisms for mild mitochondrial dysfunction alter mtDNA gene expression at a proteomic and/or translational level. Understanding these mechanisms will facilitate the development of therapeutics for mitochondrial disorders.

Epigenetic programs controlling development of the female reproductive tract (FRT) are influenced by the effects of naturally occurring bioactive agents on patterns of gene expression in FRT tissues during organizationally critical periods of foetal and perinatal life. Aberrations in such important cellular and molecular events, as may occur with exposure to natural or manmade steroid or peptide receptor-modulating agents, disrupt the developmental program and can change the developmental trajectory of FRT tissues, including the endometrium, with lasting consequences. In the pig, as in other mammals, maternal programming of FRT development begins pre-natally and is completed post-natally, when maternal effects on development can be communicated via signals transmitted in milk. Studies involving relaxin (RLX), a prototypic milk-borne morphoregulatory factor (MbF), serve as the basis for ongoing efforts to identify maternal programming events that affect uterine and cervical tissues in the neonatal pig. Data support the lactocrine hypothesis for delivery of MbFs to neonates as a specific consequence of nursing. Components of a maternally driven lactocrine mechanism for RLX-mediated signalling in neonatal FRT tissues, including evidence that milk-borne RLX is delivered into the neonatal circulation where it can act on RLX receptor (RXFP1) -positive neonatal tissues to affect their development, are in place in the pig. The fact that all newborn mammals drink milk extends the timeframe of maternal influence on neonatal development across many species. Thus, lactocrine transmission of milk-borne developmental signals is an element of the maternal epigenetic programming equation that deserves further study.

Exposure to testosterone (T) during d 30-90 of fetal life results in low-birth-weight offspring, hypergonadotropism, multifollicular ovaries, and early cessation of cyclicity. The multifollicular phenotype may result from failure of follicles to regress and consequent follicular persistence or, alternatively, increased follicular recruitment. We tested the hypothesis that prenatal exposure to excess T causes intrauterine growth retardation and increases ovarian follicular recruitment. Time-mated pregnant ewes were treated with 100 mg T propionate in cottonseed oil or vehicle twice weekly from d 30-90 of gestation. Ewes were euthanized near term, from d 139-141 of gestation (term is 147 d). After determining fetal measures and organ weights, ovaries were removed from fetuses of control and T-treated dams, and follicular distribution in each ovary was determined by morphometric quantification. Total number and percentage distribution of the various classes of follicles (primordial, primary, preantral, and antral follicles) were compared between treatment groups. Prenatally T-treated female fetuses were smaller in size, had an increased head circumference to fetal weight ratio (P < 0.01), increased adrenal to fetal weight ratio (P < 0.05), decreased number of follicles (P < 0.05), a decrease in percentage of primordial follicles (P < 0.001), and a corresponding increase in the remaining classes of follicles (P < 0.05). Ovarian findings support decreased ovarian reserve and enhanced follicular recruitment, potential contributors of early reproductive failure. The extent to which metabolic changes associated with intrauterine growth retardation contribute toward altered trajectory of ovarian folliculogenesis remains to be determined.