Pituitary adenylate cyclase activating polypeptide(PACAP) has a wide range of biological functions. We cloned the full-length c DNAs encoding PACAP and PACAP-related peptide(PRP) from the brain of largemouth bass(Micr...Pituitary adenylate cyclase activating polypeptide(PACAP) has a wide range of biological functions. We cloned the full-length c DNAs encoding PACAP and PACAP-related peptide(PRP) from the brain of largemouth bass(Micropterus salmoides) and used real-time quantitative PCR to detect PRP- PACAP m RNA expression. The PRP- PACAP cDNA has two variants expressed via alternative splicing: a long form,which encodes both PRP and PACAP,and a short form,which encodes only PACAP. Sequence analysis results are consistent with a higher conservation of PACAP than PRP peptide sequences. The expression of P ACAP-long and PACAP- short transcripts was highest in the forebrain,followed by the medulla,midbrain,pituitary,stomach,cerebellum,intestine,and kidney; however,these transcripts were either absent or were weakly expressed in the muscle,spleen,gill,heart,fatty tissue,and liver. The level of PACAP-short transcript expression was significantly higher than expression of the long transcript in the forebrain,cerebella,pituitary and intestine,but lower than that of the long transcript in the stomach. P ACAP-long and PACAP-short transcripts were first detected at the blastula stage of embryogenesis,and the level of expression increased markedly between the muscular contraction stage and 3 d post hatch(dph). The expression of PACAP- long and PACAP- short transcripts decreased significantly in the brain following 4 d fasting compared with the control diet group. The down-regulation effect was enhanced as fasting continued. Conversely,expression levels increased significantly after 3d of re-feeding. Our results suggest that PRP- PACAP acts as an important factor in appetite regulation in largemouth bass.展开更多
BACKGROUND:The effect of pituitary adenylate cyclase activating polypeptide(PACAP)during traumatic brain injury(TBI) and whether it can modulate secondary injury has not been reported previously.The present study eval...BACKGROUND:The effect of pituitary adenylate cyclase activating polypeptide(PACAP)during traumatic brain injury(TBI) and whether it can modulate secondary injury has not been reported previously.The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI.METHODS:Male Sprague Dawley rats were randomly divided into 3 treatment groups(n=6,each):sham-operated,vehicle(normal saline)^+TBI,and PACAP^+TBI.Normal saline or PACAP(1 ug/5uL) was administered intracerebroventricularly 20 minutes before TBI.Right parietal cortical contusion was produced via a weight-dropping method.Brains were extracted 24 hours after trauma.Histological changes in brains were examined by HE staining.The numbers of CD4^+ and CD8^+ T cells in blood and the spleen were detected via flow cytometry.RESULTS:In injured brain regions,edema,hemorrhage,inflammatory cell infiltration,and swollen and degenerated neurons were observed under a light microscope,and the neurons were disorderly arrayed in the hippocampi.Compared to the sham group,average CD4^+ CD8" lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI^+vehicle,and CD4^+ CD8^+ were increased.In rats administered PACAP prior to TBI,damage was attenuated as evidenced by significantly increased CD4^+,and decreased CD8^+,T lymphocytes in blood and the spleen.CONCLUSION:Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.展开更多
Pituitary adenylate cyclase-activating polypeptide（PACAP） is an endogenous peptide with neuroprotective effects on retinal neurons, but the precise mechanism underlying these effects remains unknown. Considering the...Pituitary adenylate cyclase-activating polypeptide（PACAP） is an endogenous peptide with neuroprotective effects on retinal neurons, but the precise mechanism underlying these effects remains unknown. Considering the abundance of mitochondria in retinal ganglion cells（RGCs）, we postulate that the protective effect of PACAP is associated with the regulation of mitochondrial function. RGC-5 cells were subjected to serum deprivation for 48 hours to induce apoptosis in the presence or absence of 100 nM PACAP. As revealed with the Cell Counting Kit-8 assay, PACAP at different concentrations significantly increased the viability of RGC-5 cells. PACAP also inhibited the excessive generation of reactive oxygen species in RGC-5 cells subjected to serum deprivation. We also showed by flow cytometry that PACAP inhibited serum deprivation-induced apoptosis in RGC-5 cells. The proportions of apoptotic cells and cells with mitochondria depolarization were significantly decreased with PACAP treatment. Western blot assays demonstrated that PACAP increased the levels of Bcl-2 and inhibited the compensatory increase of PAC1. Together, these data indicate protective effects of PACAP against serum deprivation-induced apoptosis in RGCs, and that the mechanism of this action is associated with maintaining mitochondrial function.展开更多
AIM:To determine the effect of pituitary adenylate cy-clase-activating polypeptide (PACAP) on left gastric artery (LGA) flow and to unveil the structural or functional important sites that may be critical for discrimi...AIM:To determine the effect of pituitary adenylate cy-clase-activating polypeptide (PACAP) on left gastric artery (LGA) flow and to unveil the structural or functional important sites that may be critical for discrimination of different receptor subtypes. METHODS: Peptides, including PACAP-27, PACAP-38, amino acid substituted PACAP-27 and C-terminus truncated analogues PACAP (27-38), were synthesized by a simultaneous multiple solid-phase peptide synthesizer. Flow probes of an ultrasound transit-time blood flowmeter were placed around the LGA of beagle dogs. Whenpeptides were infused intravenously, the blood flow was measured.RESULTS: [Ala4, Val5]-PACAP-27 caused a concentration-dependent vasodepressor action which was similar to that caused by PACAP-27. The LGA blood flow response to [Ala4, Val5]-PACAP-27 was significantly higher than that to PACAP-27, which was similar to that to vasoactive intestinal polypeptide (VIP) at the same dose. [Ala6]-PACAP-27 did not increase the peak LGA ? ow. [Gly8]-PACAP-27 showed a similar activity to VIP. [Asn24, Ser25, Ile26]-PACAP-27 did not change the activity of peptides at all doses. CONCLUSION: NH2 terminus is more important to biological activity of peptides and specifi c receptor recognition than COOH-terminus.展开更多
Pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) have been known for decades to mediate neuroendocrine and vasodilative actions via G-protein-coupled receptors of Clas...Pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) have been known for decades to mediate neuroendocrine and vasodilative actions via G-protein-coupled receptors of Class B. These are targets of imaging probes for positron emission tomography (PET) or single photon emission tomography (SPECT) in tumor diagnostics and tumor grading. However, they play only a subordinate role in the development of tracers for brain imaging. Difficulties in development of non-peptide ligands typical for cerebral receptors of PACAP and VIP are shared by all members of Class B receptor family. Essential landmarks have been confirmed for understanding of structural details of Class B receptor molecular signalling during the last five years. High relevance in the explanation of problems in ligand development for these receptors is admitted to the large N-terminal?ectodomain markedly different from Class A receptor binding sites and poorly suitable as orthosteric binding sites for the most small-molecule compounds. The present study is focused on the recently available receptor ligands for PAC1, VPAC1 and VPAC2 receptors as well as potential small-molecule lead structures suitable for use in PET or SPECT. Recently, biaryl, cyanothiophene and pentanamide structures with affinities in nM-range have been proposed as non-peptide ligands at VPAC1 and VPAC2 receptors. However, most of these ligands have been classified as non-competitive related to the orthosteric binding site of endogenous peptide ligands of VPAC receptors. For PAC1 receptors have been identified hydrazide compounds for which an inhibitory and potentially competitive mechanism of receptor binding has been postulated based on molecular docking studies.展开更多
基金Supported by the National Natural Science Foundation of China(No.31201985)the National Key Technology R&D Program of China(No.2012BAD26B03)
文摘Pituitary adenylate cyclase activating polypeptide(PACAP) has a wide range of biological functions. We cloned the full-length c DNAs encoding PACAP and PACAP-related peptide(PRP) from the brain of largemouth bass(Micropterus salmoides) and used real-time quantitative PCR to detect PRP- PACAP m RNA expression. The PRP- PACAP cDNA has two variants expressed via alternative splicing: a long form,which encodes both PRP and PACAP,and a short form,which encodes only PACAP. Sequence analysis results are consistent with a higher conservation of PACAP than PRP peptide sequences. The expression of P ACAP-long and PACAP- short transcripts was highest in the forebrain,followed by the medulla,midbrain,pituitary,stomach,cerebellum,intestine,and kidney; however,these transcripts were either absent or were weakly expressed in the muscle,spleen,gill,heart,fatty tissue,and liver. The level of PACAP-short transcript expression was significantly higher than expression of the long transcript in the forebrain,cerebella,pituitary and intestine,but lower than that of the long transcript in the stomach. P ACAP-long and PACAP-short transcripts were first detected at the blastula stage of embryogenesis,and the level of expression increased markedly between the muscular contraction stage and 3 d post hatch(dph). The expression of PACAP- long and PACAP- short transcripts decreased significantly in the brain following 4 d fasting compared with the control diet group. The down-regulation effect was enhanced as fasting continued. Conversely,expression levels increased significantly after 3d of re-feeding. Our results suggest that PRP- PACAP acts as an important factor in appetite regulation in largemouth bass.
文摘BACKGROUND:The effect of pituitary adenylate cyclase activating polypeptide(PACAP)during traumatic brain injury(TBI) and whether it can modulate secondary injury has not been reported previously.The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI.METHODS:Male Sprague Dawley rats were randomly divided into 3 treatment groups(n=6,each):sham-operated,vehicle(normal saline)^+TBI,and PACAP^+TBI.Normal saline or PACAP(1 ug/5uL) was administered intracerebroventricularly 20 minutes before TBI.Right parietal cortical contusion was produced via a weight-dropping method.Brains were extracted 24 hours after trauma.Histological changes in brains were examined by HE staining.The numbers of CD4^+ and CD8^+ T cells in blood and the spleen were detected via flow cytometry.RESULTS:In injured brain regions,edema,hemorrhage,inflammatory cell infiltration,and swollen and degenerated neurons were observed under a light microscope,and the neurons were disorderly arrayed in the hippocampi.Compared to the sham group,average CD4^+ CD8" lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI^+vehicle,and CD4^+ CD8^+ were increased.In rats administered PACAP prior to TBI,damage was attenuated as evidenced by significantly increased CD4^+,and decreased CD8^+,T lymphocytes in blood and the spleen.CONCLUSION:Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.
基金supported by grants from the Medical Scientific Research Foundation of Guangdong Province of China,No.A2016271the Natural Science Foundation of Guangdong Province of China,No.2016A030313208the Science and Technology Planning Project of Guangdong Province of China,No.2014A020212393
文摘Pituitary adenylate cyclase-activating polypeptide（PACAP） is an endogenous peptide with neuroprotective effects on retinal neurons, but the precise mechanism underlying these effects remains unknown. Considering the abundance of mitochondria in retinal ganglion cells（RGCs）, we postulate that the protective effect of PACAP is associated with the regulation of mitochondrial function. RGC-5 cells were subjected to serum deprivation for 48 hours to induce apoptosis in the presence or absence of 100 nM PACAP. As revealed with the Cell Counting Kit-8 assay, PACAP at different concentrations significantly increased the viability of RGC-5 cells. PACAP also inhibited the excessive generation of reactive oxygen species in RGC-5 cells subjected to serum deprivation. We also showed by flow cytometry that PACAP inhibited serum deprivation-induced apoptosis in RGC-5 cells. The proportions of apoptotic cells and cells with mitochondria depolarization were significantly decreased with PACAP treatment. Western blot assays demonstrated that PACAP increased the levels of Bcl-2 and inhibited the compensatory increase of PAC1. Together, these data indicate protective effects of PACAP against serum deprivation-induced apoptosis in RGCs, and that the mechanism of this action is associated with maintaining mitochondrial function.
基金Supported by (in part) Grants from Ministry of Education,Culture,Science,and Technology,Japan Society for the Promotion of Science and Special Fund of Six-Talented Peak of Jiangsu Province,No.07-B-15 (IB07)
文摘AIM:To determine the effect of pituitary adenylate cy-clase-activating polypeptide (PACAP) on left gastric artery (LGA) flow and to unveil the structural or functional important sites that may be critical for discrimination of different receptor subtypes. METHODS: Peptides, including PACAP-27, PACAP-38, amino acid substituted PACAP-27 and C-terminus truncated analogues PACAP (27-38), were synthesized by a simultaneous multiple solid-phase peptide synthesizer. Flow probes of an ultrasound transit-time blood flowmeter were placed around the LGA of beagle dogs. Whenpeptides were infused intravenously, the blood flow was measured.RESULTS: [Ala4, Val5]-PACAP-27 caused a concentration-dependent vasodepressor action which was similar to that caused by PACAP-27. The LGA blood flow response to [Ala4, Val5]-PACAP-27 was significantly higher than that to PACAP-27, which was similar to that to vasoactive intestinal polypeptide (VIP) at the same dose. [Ala6]-PACAP-27 did not increase the peak LGA ? ow. [Gly8]-PACAP-27 showed a similar activity to VIP. [Asn24, Ser25, Ile26]-PACAP-27 did not change the activity of peptides at all doses. CONCLUSION: NH2 terminus is more important to biological activity of peptides and specifi c receptor recognition than COOH-terminus.
文摘Pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) have been known for decades to mediate neuroendocrine and vasodilative actions via G-protein-coupled receptors of Class B. These are targets of imaging probes for positron emission tomography (PET) or single photon emission tomography (SPECT) in tumor diagnostics and tumor grading. However, they play only a subordinate role in the development of tracers for brain imaging. Difficulties in development of non-peptide ligands typical for cerebral receptors of PACAP and VIP are shared by all members of Class B receptor family. Essential landmarks have been confirmed for understanding of structural details of Class B receptor molecular signalling during the last five years. High relevance in the explanation of problems in ligand development for these receptors is admitted to the large N-terminal?ectodomain markedly different from Class A receptor binding sites and poorly suitable as orthosteric binding sites for the most small-molecule compounds. The present study is focused on the recently available receptor ligands for PAC1, VPAC1 and VPAC2 receptors as well as potential small-molecule lead structures suitable for use in PET or SPECT. Recently, biaryl, cyanothiophene and pentanamide structures with affinities in nM-range have been proposed as non-peptide ligands at VPAC1 and VPAC2 receptors. However, most of these ligands have been classified as non-competitive related to the orthosteric binding site of endogenous peptide ligands of VPAC receptors. For PAC1 receptors have been identified hydrazide compounds for which an inhibitory and potentially competitive mechanism of receptor binding has been postulated based on molecular docking studies.
