The direct effects of recombinant porcine leptin on porcine granulosa cells were studied to test the hypothesis that leptin, acting through the nuclear transcription factor signal transducer and activator of transcription 3 (STAT-3), modulates sterol regulatory element-binding protein 1 (SREBP1) thereby increasing steroidogenesis. In porcine granulosa cells in culture over 48 h, leptin at 10 ng/ml increased progesterone accumulation 3-fold while it was reduced by leptin at 1000 ng/ml. Leptin had no effect on progression of granulosa cells through the cell cycle nor on the frequency of cell death. Leptin treatment at 24 or 48 h of culture resulted in dose-dependent 2- to 4-fold increases in tyrosine phosphorylation of STAT-3. Leptin had a biphasic effect on the abundance of membrane-bound and transcriptionally active forms of SREBP1. In transient transfection of primary porcine granulosa cells, the plasmid expressing the transcriptionally active form of SREPB-1 induced transcription of the key regulator of steroidogenesis, the steroidogenic acute regulatory protein (StAR). StAR transcription was also increased by the low dose of leptin and was further upregulated in the presence of the SREBP plasmid. Leptin at 1000 ng/ml inhibited SREBP1-induced StAR expression. Thus, leptin, acting through STAT-3, modulates steroidogenesis in a biphasic and dose-dependent manner, and SREBP1 induction of StAR expression may be in the cascade of regulatory events.

Weaning is associated with increased occurrence of infections and diseases in piglets. Recent findings indicate that weaning induces mitochondrial dysfunction and oxidative stress conditions that more severely impact smaller piglets. The objective of this study was to characterize the molecular mechanisms underlying these physiological consequences and the relation with systemic inflammatory status in both normal and low birth weight (NBW and LBW) piglets throughout the peri-weaning period. To conduct the study, 30 sows were inseminated, and specific piglets from their litters were assigned to one of two experimental groups: NBW (n = 60, 1.73 ± 0.01 kg,) and LBW piglets weighing less than 1.2 kg (n = 60, 1.01 ± 0.01 kg). Then, 10 piglets from each group were selected at 14, 21 (weaning), 23, 25, 29 and 35 days of age to collect organ and plasma samples. Specific porcine RT2 Profiler™ PCR Arrays related to mitochondrial function, oxidative stress, inflammation and apoptosis processes were first used to target genes that are modulated after weaning in NBW piglets (d 23 and d 35 vs. d 14). Expression of selected genes was evaluated by quantitative PCR. These analyses revealed that expression of inflammatory genes CXCL10 and CCL19 increased after weaning in intestinal mucosa, while expression of genes encoding subunits of the mitochondrial respiratory chain was downregulated in liver and kidney of both groups. Interestingly, major modulators of mitophagy (BNIP3), cell survival (BCL2A1) and antioxidant defense system (TXNRD2, GPx3, HMOX1) were found to be highly expressed in NBW piglets. The systemic levels of TNF-α and IL1-β significantly increased following weaning and were higher in NBW piglets. These results provide novel information about the molecular origin of mitochondrial dysfunction and oxidative stress observed in weaned piglets and suggest that clearance of dysfunctional mitochondria, antioxidant defenses and inflammatory response are compromised in LBW piglets.

The impact of diet deprivation and subsequent over-allowance on the metabolite status, mammary development, and mammary gene expression in prepubertal gilts was determined. Forty-seven gilts were reared under a conventional (control, CTL; n = 23) or an experimental (treatment, TRT; n = 24) dietary regimen. The later regimen (consisting of diet deprivation and subsequent over-allowance) provided 70 (restriction diet, RES) and 115% (over-allowance diet, OVER) of the protein and DE contents provided by the CTL diet. Experimental diets were fed ad libitum starting at 27.7 ± 3.4 kg of BW as follows: 3 wk RES, 3 wk OVER, 4 wk RES, and 4 wk OVER. At each diet change, BW and individual feed intakes were measured, and blood samples for metabolite and IGF-I assays were obtained. Some gilts (11 CTL and 12 TRT) were slaughtered on d 235 (after reaching puberty) to collect mammary tissue for compositional analyses and measures of gene expression. Body weight gain (P < 0.01) and G:F (P < 0.05) of gilts were reduced during each period with the RES diet; however, there was no compensatory growth during the periods when the OVER diet was fed. Feeding the RES diet reduced concentrations of urea and IGF-I (P < 0.01) and feeding the OVER diet increased FFA (P < 0.01) and glucose (P < 0.10) in TRT gilts compared with CTL gilts. The TRT gilts had less parenchymal tissue (P < 0.05) and tended to have less total parenchymal fat and protein (P < 0.10) than CTL gilts. The mammary mRNA relative abundance of the signal transducers and activators of transduction 5B was decreased in TRT compared with CTL gilts (P < 0.05). In conclusion, the diet deprivation and over-allowance regimen used in the growing-finishing period did not have beneficial effects on mammary development after puberty. In fact, a detrimental effect was observed.

The impacts of diet deprivation and subsequent overallowance during gestation on mammary gene expression and development and lactation performance were determined. Gilts were reared under a conventional (control, CTL; n = 59) or an experimental (treatment, TRT; n = 56) dietary regimen during gestation. The experimental regimen provided 70% (restriction diet, RES) and 115% (overallowance diet, OVER) of the protein and DE contents provided by the CTL diet. The RES diet was given during the first 10 wk of gestation followed by the OVER diet until farrowing. Some gilts (14 CTL and 14 TRT) were slaughtered on d 110 of gestation, and the others were allowed to farrow. Of these remaining sows, 28 (14 CTL and 14 TRT) were slaughtered on d 21 of lactation, and the rest underwent a second lactation. At each slaughter, mammary tissue was collected for compositional analyses and assessment of gene expression. Milk samples were collected on d 17 of the first lactation. Litter size was standardized to 11 ± 1, and piglets were weighed weekly until d 18 in both parities. The BW and back fat thickness of TRT first-parity sows were less than those of CTL sows in gestation (P< 0.05), and their BW was also less in lactation (P < 0.05). The BW of TRT second-parity sows was still less at mating (P < 0.05) and tended to be less on d 1 of lactation (P < 0.10) compared with CTL sows. There were no differences in piglet growth between CTL and TRT litters in either parity, yet mammary development and mammary gene expression were affected by treatment. There was less parenchymal tissue (P < 0.01) at the end of the first gestation in TRT than in CTL sows, but parenchymal tissue composition was not altered by treatment. Relative abundance of IGF-1 (P < 0.05), ornithine decarboxylase (P < 0.05), signal transducer and activator of transcription 5B (P < 0.05), and whey acidic protein (WAP, P < 0.01) genes in parenchyma at the end of the first gestation was lower in TRT than in CTL sows, and the effect on WAP genes was still present at the end of the first lactation (P < 0.01). Mammary composition at the end of the first lactation and milk composition were unaffected by treatment. In conclusion, feed deprivation and subsequent overallowance in gestation had unfavorable effects on sow BW, back fat, mammary development, and mammary gene expression at the end of gestation, but piglet growth rate over the 2 parities was not affected.