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Changes in the cholinergic innervation pattern of porcine ovaries with cysts induced by dexamethasone administration.

Kozłowska A, Majewski M, Jana B - J. Mol. Neurosci. (2014)

Bottom Line: We revealed earlier that induction of ovarian cysts in gilts by dexamethasone phosphate disodium salt (DXM) administration from the follicular phase of the estrous cycle (EC) changed the cholinergic innervation of the gonad.In the cystic ovaries, VAChT-, nNOS- and SOM-IR fibres were found around cysts and small tertiary follicles; nNOS-IR and also VAChT-IR fibres were observed near secondary follicles and veins; and VAChT- and nNOS-IR fibres were not found around cortical arteries.The number of VIP-IR fibres increased near the cysts and within the ground plexus, while the number of VAChT-IR fibres decreased within the medullar part of this structure.

View Article: PubMed Central - PubMed

Affiliation: Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima 10, 10-748, Olsztyn, Poland, kozlowska.anna@uwm.edu.pl.

ABSTRACT
We revealed earlier that induction of ovarian cysts in gilts by dexamethasone phosphate disodium salt (DXM) administration from the follicular phase of the estrous cycle (EC) changed the cholinergic innervation of the gonad. In the present study, the innervation of porcine ovaries by vesicular acetylcholine transporter (VAChT)-, neuronal nitric oxide synthase (nNOS)-, vasoactive intestinal peptide (VIP)- and somatostatin (SOM)-immunoreactive (IR) fibres, after induction of cystic changes from the middle luteal phase of the EC, was determined. The cystic changes were induced by DXM injections from days 7 to 21 of the EC, and 11 days later, the ovaries were collected. In the cystic ovaries, VAChT-, nNOS- and SOM-IR fibres were found around cysts and small tertiary follicles; nNOS-IR and also VAChT-IR fibres were observed near secondary follicles and veins; and VAChT- and nNOS-IR fibres were not found around cortical arteries. The number of VIP-IR fibres increased near the cysts and within the ground plexus, while the number of VAChT-IR fibres decreased within the medullar part of this structure. Thus, our study showed changes in the cholinergic innervation pattern of the porcine cystic ovaries induced from the middle phase of the cycle and confirmed that cystic ovary innervation depends partly on the phase of the EC in which the induction of cysts was started.

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Immunohistochemical localization of VIP-IR nerve fibres in the cortical part of the ovary of the control (CON) and DXM-treated (DXM) gilts. An increase in the number of VIP-positive nerve fibres, formed bunches, in the area of the ground plexus (13) as well as near the cyst (19) in the ovary of the DXM-treated animal compared to the CON group (12 and 18, respectively). A lack of VIP-IR nerve fibres around the primordial (15) and secondary (17) follicles as well as the vein (21) after DXM administration and their presence in the CON group (14, 16 and 20, respectively). Negative control for VIP in the ovary of the CON gilt (22). Arrows, nerve fibres; PF primordial follicle, SF secondary follicle, TF 4–6 tertiary follicle 4–6 mm in diameter, C cyst, V vein. Scale bar 25 μm
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Fig3: Immunohistochemical localization of VIP-IR nerve fibres in the cortical part of the ovary of the control (CON) and DXM-treated (DXM) gilts. An increase in the number of VIP-positive nerve fibres, formed bunches, in the area of the ground plexus (13) as well as near the cyst (19) in the ovary of the DXM-treated animal compared to the CON group (12 and 18, respectively). A lack of VIP-IR nerve fibres around the primordial (15) and secondary (17) follicles as well as the vein (21) after DXM administration and their presence in the CON group (14, 16 and 20, respectively). Negative control for VIP in the ovary of the CON gilt (22). Arrows, nerve fibres; PF primordial follicle, SF secondary follicle, TF 4–6 tertiary follicle 4–6 mm in diameter, C cyst, V vein. Scale bar 25 μm

Mentions: Compared to the tertiary follicles measuring 4–6 mm in the CON group (Figs. 5a—VAChT, 5b—nNOS and 27—SOM), the presence of nerve fibres storing VAChT (Fig. 6a), nNOS (Fig. 6b) and SOM (Fig. 28) around cysts was revealed. In the ovaries of DXM-treated gilts, in contrast to the CON group (Figs. 1a—VAChT, 1b—nNOS, 3a—VAChT, 3b—nNOS and 25—SOM), VAChT- and nNOS-IR fibres were found near the secondary (Fig. 2a, b; respectively) and tertiary follicles measuring up to 3 mm (Fig. 4a, b; respectively), while SOM-IR fibres were visible within the ground plexus (Fig. 26) and around the tertiary follicles measuring up to 3 mm. Furthermore, the populations of the VIP-IR nerve fibres created thick fascicles and more were found within the ground plexus (P < 0.001; Fig. 13) and in the vicinity of the cysts (P < 0.05; Fig. 19) after DXM administration than in the CON group (Figs. 12 and 18, respectively). Neither VAChT- and nNOS-IR nerves supplying arteries nor VIP-IR terminals supplying primordial (Fig. 15) and secondary follicles (Fig. 17) and veins (Fig. 21) were found in the cystic ovaries, when compared to the CON group (Figs. 14, 16 and 20, respectively). Application of DXM did not significantly change the population of VAChT- or nNOS-IR nerve fibres within the ground plexus as well as the number of VAChT-IR fibres around the veins. The density of VIP-containing fibres near the tertiary follicles (measuring to 3 mm in diameter) and arteries as well as VAChT, nNOS, VIP and SOM supplying the interstitial gland was similar in the CON and DXM groups. CLs, present only in the control ovaries, were sporadically innervated by single VAChT-IR nerve fibres. No immunostaining for VAChT (Fig. 7a), nNOS (Fig. 7b), VIP (Fig. 22) and SOM (Fig. 29) was detected in the control ovaries when the primary antibodies were substituted with normal IgGs (in relation to VAChT—rabbit, nNOS and VIP—mouse and SOM—rat).Figs. 1–7


Changes in the cholinergic innervation pattern of porcine ovaries with cysts induced by dexamethasone administration.

Kozłowska A, Majewski M, Jana B - J. Mol. Neurosci. (2014)

Immunohistochemical localization of VIP-IR nerve fibres in the cortical part of the ovary of the control (CON) and DXM-treated (DXM) gilts. An increase in the number of VIP-positive nerve fibres, formed bunches, in the area of the ground plexus (13) as well as near the cyst (19) in the ovary of the DXM-treated animal compared to the CON group (12 and 18, respectively). A lack of VIP-IR nerve fibres around the primordial (15) and secondary (17) follicles as well as the vein (21) after DXM administration and their presence in the CON group (14, 16 and 20, respectively). Negative control for VIP in the ovary of the CON gilt (22). Arrows, nerve fibres; PF primordial follicle, SF secondary follicle, TF 4–6 tertiary follicle 4–6 mm in diameter, C cyst, V vein. Scale bar 25 μm
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Related In: Results  -  Collection

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Fig3: Immunohistochemical localization of VIP-IR nerve fibres in the cortical part of the ovary of the control (CON) and DXM-treated (DXM) gilts. An increase in the number of VIP-positive nerve fibres, formed bunches, in the area of the ground plexus (13) as well as near the cyst (19) in the ovary of the DXM-treated animal compared to the CON group (12 and 18, respectively). A lack of VIP-IR nerve fibres around the primordial (15) and secondary (17) follicles as well as the vein (21) after DXM administration and their presence in the CON group (14, 16 and 20, respectively). Negative control for VIP in the ovary of the CON gilt (22). Arrows, nerve fibres; PF primordial follicle, SF secondary follicle, TF 4–6 tertiary follicle 4–6 mm in diameter, C cyst, V vein. Scale bar 25 μm
Mentions: Compared to the tertiary follicles measuring 4–6 mm in the CON group (Figs. 5a—VAChT, 5b—nNOS and 27—SOM), the presence of nerve fibres storing VAChT (Fig. 6a), nNOS (Fig. 6b) and SOM (Fig. 28) around cysts was revealed. In the ovaries of DXM-treated gilts, in contrast to the CON group (Figs. 1a—VAChT, 1b—nNOS, 3a—VAChT, 3b—nNOS and 25—SOM), VAChT- and nNOS-IR fibres were found near the secondary (Fig. 2a, b; respectively) and tertiary follicles measuring up to 3 mm (Fig. 4a, b; respectively), while SOM-IR fibres were visible within the ground plexus (Fig. 26) and around the tertiary follicles measuring up to 3 mm. Furthermore, the populations of the VIP-IR nerve fibres created thick fascicles and more were found within the ground plexus (P < 0.001; Fig. 13) and in the vicinity of the cysts (P < 0.05; Fig. 19) after DXM administration than in the CON group (Figs. 12 and 18, respectively). Neither VAChT- and nNOS-IR nerves supplying arteries nor VIP-IR terminals supplying primordial (Fig. 15) and secondary follicles (Fig. 17) and veins (Fig. 21) were found in the cystic ovaries, when compared to the CON group (Figs. 14, 16 and 20, respectively). Application of DXM did not significantly change the population of VAChT- or nNOS-IR nerve fibres within the ground plexus as well as the number of VAChT-IR fibres around the veins. The density of VIP-containing fibres near the tertiary follicles (measuring to 3 mm in diameter) and arteries as well as VAChT, nNOS, VIP and SOM supplying the interstitial gland was similar in the CON and DXM groups. CLs, present only in the control ovaries, were sporadically innervated by single VAChT-IR nerve fibres. No immunostaining for VAChT (Fig. 7a), nNOS (Fig. 7b), VIP (Fig. 22) and SOM (Fig. 29) was detected in the control ovaries when the primary antibodies were substituted with normal IgGs (in relation to VAChT—rabbit, nNOS and VIP—mouse and SOM—rat).Figs. 1–7

Bottom Line: We revealed earlier that induction of ovarian cysts in gilts by dexamethasone phosphate disodium salt (DXM) administration from the follicular phase of the estrous cycle (EC) changed the cholinergic innervation of the gonad.In the cystic ovaries, VAChT-, nNOS- and SOM-IR fibres were found around cysts and small tertiary follicles; nNOS-IR and also VAChT-IR fibres were observed near secondary follicles and veins; and VAChT- and nNOS-IR fibres were not found around cortical arteries.The number of VIP-IR fibres increased near the cysts and within the ground plexus, while the number of VAChT-IR fibres decreased within the medullar part of this structure.

View Article: PubMed Central - PubMed

Affiliation: Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima 10, 10-748, Olsztyn, Poland, kozlowska.anna@uwm.edu.pl.

ABSTRACT
We revealed earlier that induction of ovarian cysts in gilts by dexamethasone phosphate disodium salt (DXM) administration from the follicular phase of the estrous cycle (EC) changed the cholinergic innervation of the gonad. In the present study, the innervation of porcine ovaries by vesicular acetylcholine transporter (VAChT)-, neuronal nitric oxide synthase (nNOS)-, vasoactive intestinal peptide (VIP)- and somatostatin (SOM)-immunoreactive (IR) fibres, after induction of cystic changes from the middle luteal phase of the EC, was determined. The cystic changes were induced by DXM injections from days 7 to 21 of the EC, and 11 days later, the ovaries were collected. In the cystic ovaries, VAChT-, nNOS- and SOM-IR fibres were found around cysts and small tertiary follicles; nNOS-IR and also VAChT-IR fibres were observed near secondary follicles and veins; and VAChT- and nNOS-IR fibres were not found around cortical arteries. The number of VIP-IR fibres increased near the cysts and within the ground plexus, while the number of VAChT-IR fibres decreased within the medullar part of this structure. Thus, our study showed changes in the cholinergic innervation pattern of the porcine cystic ovaries induced from the middle phase of the cycle and confirmed that cystic ovary innervation depends partly on the phase of the EC in which the induction of cysts was started.

Show MeSH
Related in: MedlinePlus