Expression of proopiomelanocortin, proenkephalin and prodynorphin genes in porcine luteal cells

Abstract

The objective of the study was to examine the expression of the genes coding for proopiomelanocortin (POMC), proenkephalin (PENK) and prodynorphin (PDYN) in porcine luteal cells isolated from corpora lutea (CL) collected on days 3-6, 8-10 and 13-16 of the oestrous cycle. Total RNA was purified from nonincubated cells and from cells incubated for 48 h in the absence or presence of luteinising hormone (LH). The semi-quantitative RT-PCR technique, involving coamplification of the target and control cDNA (β-actin or 18S rRNA), was used to examine gene expression. It was found that the genes coding for opioid precursors are expressed in both non-incubated and incubated porcine luteal cells representing the early, mid-and late luteal phase. In non-incubated cells, only POMC mRNA content changed during CL development, whereas the expression of PENK and PDYN genes remained relatively constant. Additionally, the treatment of cells with LH markedly affected the expression of POMC and PENK, but no influence on PDYN expression was observed. The present study indicates that porcine luteal cells may produce opioid peptides and that gene expression of their precursors (except for PDYN) may be modulated in these cells by LH. Moreover, the present results support the involvement of opioid peptides in local regulation within the CL of the pig.

Peri‐conceptional under‐nutrition alters transcriptomic profile in the endometrium during the peri‐implantation period—The study in domestic pigs

“…Primer specificities were confirmed in silico with Basic Local Alignment Search Tool (BLAST). Gene expression levels were calculated using the ΔΔ Ct method and normalized using the geometric mean of the expression levels of three reference genes— glyceraldehyde 3‐phosphate dehydrogenase ( GAPDH ; Bogacka, Przała, Siawrys, Kamiński, & Smolińska, ), β ‐actin ( ACTB ; Staszkiewicz et al., ), and 18S ribosomal RNA ( 18S rRNA; Franczak, Zgleic, et al., ) were used.…”

Transcriptomic analysis of the myometrium during peri-implantation period and luteolysis–the study on the pig model

“…Specific primers (Table 1 ) were designed using the Primer Express 3.0 software (Life Technologies), and primer specificities were confirmed with Basic Local Alignment Search Tool (BLAST). Gene expression levels were calculated using the ΔΔCt method and normalised using the geometric mean of the expression levels of two reference genes—glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ; Bogacka et al 2006 ) and β-actin ( ACTB ; Staszkiewicz et al 2007 ). The significant difference in gene expression between the myometrium of pigs during the implantation and luteolytic periods was analysed via Student’s t test.…”

Peri‐conceptional under‐nutrition alters the expression ofTRIM28 andZFP57 in the endometrium and embryos during peri‐implantation period in domestic pigs

“…Non‐template controls were used for each set of primers to confirm reaction specificity. The relative expression of TRIM28 and ZFP57 was calculated using the ΔΔCt method and normalized using the geometric mean of expression levels of three reference genes (Table ): β‐actin (ACTB; Staszkiewicz et al., ), glyceraldehyde 3‐phosphate dehydrogenase (GAPDH; Bogacka, Przała, Siawrys, Kaminski, & Smolinska, ) and 18S rRNA (Franczak, Zglejc et al., ).…”

“…: β-actin (ACTB;Staszkiewicz et al, 2007), glyceraldehyde 3-phosphate dehydrogenase (GAPDH;Bogacka, Przała, Siawrys, Kaminski, & Smolinska, 2006) and 18S rRNA.…”