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Effect of chronic administration of morphine on the expression of BDNF mRNA in the rat lumbar spinal cord
S284
Abstracts
Discussion: Both MTHFR 677 T and 1298 C alleles increased the susceptibility to the onset and progression of DN in Iranians with T2DM. Keywords: MTHFR C677T, MTHFR A1298C, Microalbuminuria, Macroalbuminuria, Type 2 diabetes mellitus doi:10.1016/j.clinbiochem.2011.08.706
Poster – [A-10-493-8] The concomitant presence of eNOS 894 T and ACE D alleles is associated with diabetic nephropathy and its progression Rahimi Zohreha, Asad Vaisi-Rayganib, Ziba Rahimib a Medical School, Daneshgah Avenue, Kermanshah, Kermanshah, Iran b Medical School, Daneshgah Avenue, Kermanshah, Iran E-mail address: [email protected] (R. Zohreh) Introduction: The present study investigated the influence of insertion (I)/deletion (D) polymorphism of the angiotensin II– converting enzyme (ACE) gene in combination with endothelial nitric oxide (eNOS) G894T polymorphism on the predisposition to diabetic nephropathy (DN) and its progression. Materials and methods: Using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) method the ACE and eNOS polymorphisms were detected in 72 microalbuminuric, 68 macroalbuminuric and 72 normoalbuinuric type 2 diabetes mellitus (T2DM) patients from West of Iran. Results: The presence of eNOS T and ACE D alleles separately increased the risk of macroalbuminuria (1.36-fold, p = 0.27 and 1.6-fold, p = 0.062, respectively) and progression of DN (1.31-fold, p = 0.34 and 1.3-fold, p = 0.036). However, the combined presence of both alleles was associated with higher risk of macroalbuminuria (5-fold, p = 0.05) and progression from micro-to macro-albuminuria (3.33-fold, p = 0.18). Also, while the presence of ACE D or eNOS T alleles alone did not increase or had a little effect on the risk of microalbuminuria (OR = 0.97 and OR= 1.04, respectively) the combined presence of both alleles increased the risk of microalbuminuria 1.5 times. Discussion: Our study indicated that the concomitant presence of both ACE D and eNOS T alleles is a risk factor for microalbuminuria and increased the risk of macroalbuminuria and progression of DN compared to the existence of each polymorphism alone which could be attributed to the increasing activity of ACE and angiotensin II level in the presence of D allele and decreasing NO production in the presence of T allele accelerating diabetic nephropathy and its progression. Keywords: ACE I/D, eNOS G894T, Microalbuminuria, Macroalbuminuria, Type 2 diabetes mellitus doi:10.1016/j.clinbiochem.2011.08.707
E.poster – [A-10-499-2] Expression levels of BDNF gene and epigenetic modifications of its promoters in the ventral tegmental area of morphine addicted rats Farideh Jalali Mashayekhi, Mozhgan Rasti, Mostafa Rahvar, Pooneh Mokaram, Mohammad Reza Namavar, Ali Akbar Owji Shiraz University of Medical Sciences, Shiraz, Iran E-mail address: [email protected] (A.A. Owji)
neurotrophic factor (BDNF) plays a role in mediating molecular, cellular, and behavioral adaptations underlying drug addiction. Here, we examined the influence of chronic morphine treatment on the expression of BDNF mRNA in the ventral tegmental area (VTA), a major brain reward region. We also studied whether alternations in BDNF mRNA levels within the VTA are associated with levels of H4 acetylation, H3 acetylation and histone methylation around three promoters of the BDNF gene. Methods: VTA of rats was subjected to quantitative RT-PCR to analyze the expression of BDNF gene. Chromatin immunoprecipitation assays were also carried out on the same tissues to assess levels of histone acetylation and histone methylation within the promoters of the gene for BDNF. Results: VTA levels of BDNF mRNA were significantly increased (3.4 ± 0.7 fold relative to control rats) 7 days but not 2 h or 24 h after the last injection of morphine. Consistently, after 7 days of morphine withdrawal, levels of histone methylation at the BDNF gene promoter of rats treated by chronic morphine were lower than control animals. Morphine withdrawal caused no changes in the levels of histone acetylation at any of the promoters of BDNF gene of rats treated by chronic morphine. Conclusion: These data provide the first in vivo demonstration of the involvement of chromatin remodeling in morphine-induced regulation of gene expression in the VTA of rats. Keywords: Chromatin remodeling, BDNF, Morphine, Rat doi:10.1016/j.clinbiochem.2011.08.708
Poster – [A-10-499-3] Effect of chronic administration of morphine on the expression of BDNF mRNA in the rat lumbar spinal cord Zahra Khoshdel, Mohammad Ali Takhshid, Farideh Jalali Mashayekhi, Mostafa Rahvar, Ebrahim Eftekhar, Ali Akbar Owji Shiraz University of Medical Sciences, Shiraz, Iran E-mail address: [email protected] (A.A. Owji) Introduction: Brain derived neurotrophic factor (BDNF) is a neurotrophin expressed in several areas of the central nervous system. The BDNF immunoreactivity that has been detected in laminae I and II of the rat spinal cord is suggested to be raised from primary nociceptive neurons in the dorsal root ganglion and, therefore, may be involved in responses to painful stimuli. As BDNF plays a role in the chronic responses produced by addictive drugs, we designed the present study to examine the expression of BDNF mRNA in both normal and morphine treated rats. Methods: Opiate addiction was induced by a progressive injection of rats with morphine. Rats were injected with morphine sulfate, 10– 50 mg/kg or saline intraperitonealy two times a day for 10 consecutive days. Lumbar spinal cord was isolated for RNA extraction 24 h and 3 weeks after the last injection of morphine or saline. Spinal cord was assayed for the expression of BDNF mRNA using RT-PCR method. Levels of BDNF mRNA were normalized to those of ß-actin. Results: Expression of BDNF mRNA in the rat spinal cord was revealed by RT PCR. Chronic injection of morphine significantly increased the levels of mRNA for BDNF in the lumbar spinal cord. Conclusions: BDNF mRNA is expressed in the lumbar spinal cord of rats. Implications of over-expression of BDNF mRNA in the development of addiction and tolerance to morphine are the subject of further investigation. Keywords: Morphine, Spinal cord, BDNF, mRNA
Introduction: Epigenetic mechanisms are suggested to make an important contribution to the persistent changes in neuronal gene expression that occur upon exposure to opiates. Brain-derived
doi:10.1016/j.clinbiochem.2011.08.709
Abstracts
Discussion: Both MTHFR 677 T and 1298 C alleles increased the susceptibility to the onset and progression of DN in Iranians with T2DM. Keywords: MTHFR C677T, MTHFR A1298C, Microalbuminuria, Macroalbuminuria, Type 2 diabetes mellitus doi:10.1016/j.clinbiochem.2011.08.706
Poster – [A-10-493-8] The concomitant presence of eNOS 894 T and ACE D alleles is associated with diabetic nephropathy and its progression Rahimi Zohreha, Asad Vaisi-Rayganib, Ziba Rahimib a Medical School, Daneshgah Avenue, Kermanshah, Kermanshah, Iran b Medical School, Daneshgah Avenue, Kermanshah, Iran E-mail address: [email protected] (R. Zohreh) Introduction: The present study investigated the influence of insertion (I)/deletion (D) polymorphism of the angiotensin II– converting enzyme (ACE) gene in combination with endothelial nitric oxide (eNOS) G894T polymorphism on the predisposition to diabetic nephropathy (DN) and its progression. Materials and methods: Using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) method the ACE and eNOS polymorphisms were detected in 72 microalbuminuric, 68 macroalbuminuric and 72 normoalbuinuric type 2 diabetes mellitus (T2DM) patients from West of Iran. Results: The presence of eNOS T and ACE D alleles separately increased the risk of macroalbuminuria (1.36-fold, p = 0.27 and 1.6-fold, p = 0.062, respectively) and progression of DN (1.31-fold, p = 0.34 and 1.3-fold, p = 0.036). However, the combined presence of both alleles was associated with higher risk of macroalbuminuria (5-fold, p = 0.05) and progression from micro-to macro-albuminuria (3.33-fold, p = 0.18). Also, while the presence of ACE D or eNOS T alleles alone did not increase or had a little effect on the risk of microalbuminuria (OR = 0.97 and OR= 1.04, respectively) the combined presence of both alleles increased the risk of microalbuminuria 1.5 times. Discussion: Our study indicated that the concomitant presence of both ACE D and eNOS T alleles is a risk factor for microalbuminuria and increased the risk of macroalbuminuria and progression of DN compared to the existence of each polymorphism alone which could be attributed to the increasing activity of ACE and angiotensin II level in the presence of D allele and decreasing NO production in the presence of T allele accelerating diabetic nephropathy and its progression. Keywords: ACE I/D, eNOS G894T, Microalbuminuria, Macroalbuminuria, Type 2 diabetes mellitus doi:10.1016/j.clinbiochem.2011.08.707
E.poster – [A-10-499-2] Expression levels of BDNF gene and epigenetic modifications of its promoters in the ventral tegmental area of morphine addicted rats Farideh Jalali Mashayekhi, Mozhgan Rasti, Mostafa Rahvar, Pooneh Mokaram, Mohammad Reza Namavar, Ali Akbar Owji Shiraz University of Medical Sciences, Shiraz, Iran E-mail address: [email protected] (A.A. Owji)
neurotrophic factor (BDNF) plays a role in mediating molecular, cellular, and behavioral adaptations underlying drug addiction. Here, we examined the influence of chronic morphine treatment on the expression of BDNF mRNA in the ventral tegmental area (VTA), a major brain reward region. We also studied whether alternations in BDNF mRNA levels within the VTA are associated with levels of H4 acetylation, H3 acetylation and histone methylation around three promoters of the BDNF gene. Methods: VTA of rats was subjected to quantitative RT-PCR to analyze the expression of BDNF gene. Chromatin immunoprecipitation assays were also carried out on the same tissues to assess levels of histone acetylation and histone methylation within the promoters of the gene for BDNF. Results: VTA levels of BDNF mRNA were significantly increased (3.4 ± 0.7 fold relative to control rats) 7 days but not 2 h or 24 h after the last injection of morphine. Consistently, after 7 days of morphine withdrawal, levels of histone methylation at the BDNF gene promoter of rats treated by chronic morphine were lower than control animals. Morphine withdrawal caused no changes in the levels of histone acetylation at any of the promoters of BDNF gene of rats treated by chronic morphine. Conclusion: These data provide the first in vivo demonstration of the involvement of chromatin remodeling in morphine-induced regulation of gene expression in the VTA of rats. Keywords: Chromatin remodeling, BDNF, Morphine, Rat doi:10.1016/j.clinbiochem.2011.08.708
Poster – [A-10-499-3] Effect of chronic administration of morphine on the expression of BDNF mRNA in the rat lumbar spinal cord Zahra Khoshdel, Mohammad Ali Takhshid, Farideh Jalali Mashayekhi, Mostafa Rahvar, Ebrahim Eftekhar, Ali Akbar Owji Shiraz University of Medical Sciences, Shiraz, Iran E-mail address: [email protected] (A.A. Owji) Introduction: Brain derived neurotrophic factor (BDNF) is a neurotrophin expressed in several areas of the central nervous system. The BDNF immunoreactivity that has been detected in laminae I and II of the rat spinal cord is suggested to be raised from primary nociceptive neurons in the dorsal root ganglion and, therefore, may be involved in responses to painful stimuli. As BDNF plays a role in the chronic responses produced by addictive drugs, we designed the present study to examine the expression of BDNF mRNA in both normal and morphine treated rats. Methods: Opiate addiction was induced by a progressive injection of rats with morphine. Rats were injected with morphine sulfate, 10– 50 mg/kg or saline intraperitonealy two times a day for 10 consecutive days. Lumbar spinal cord was isolated for RNA extraction 24 h and 3 weeks after the last injection of morphine or saline. Spinal cord was assayed for the expression of BDNF mRNA using RT-PCR method. Levels of BDNF mRNA were normalized to those of ß-actin. Results: Expression of BDNF mRNA in the rat spinal cord was revealed by RT PCR. Chronic injection of morphine significantly increased the levels of mRNA for BDNF in the lumbar spinal cord. Conclusions: BDNF mRNA is expressed in the lumbar spinal cord of rats. Implications of over-expression of BDNF mRNA in the development of addiction and tolerance to morphine are the subject of further investigation. Keywords: Morphine, Spinal cord, BDNF, mRNA
Introduction: Epigenetic mechanisms are suggested to make an important contribution to the persistent changes in neuronal gene expression that occur upon exposure to opiates. Brain-derived
doi:10.1016/j.clinbiochem.2011.08.709