RESUMO
The endangered black-footed ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25 degrees C v. 37 degrees C); (3) type of medium (Ham's F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P < 0.05) at 25 degrees C than at 37 degrees C in Ham's or TYB, but temperature did not affect (P > 0.05) acrosomal integrity. Overall, TYB maintained optimal (P < 0.05) sperm motility compared with Ham's medium, but Ham's medium maintained more (P < 0.05) intact acrosomes than TYB. Slow cooling (0.2 degrees C min(-1)) was optimal (P < 0.05) compared to rapid cooling (1 degrees C min(-1)), and ultra-rapid cooling (9 degrees C min(-1)) was found to be highly detrimental (P < 0.05). Results obtained in TYB with 0% or 4% glycerol were comparable (P > 0.05), indicating that 4% glycerol was non-toxic to ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to black-footed ferret spermatozoa is derived largely from the rate of cooling.
Assuntos
Acrossomo/fisiologia , Criopreservação/veterinária , Furões/fisiologia , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides/fisiologia , Animais , Temperatura Baixa , Conservação dos Recursos Naturais , Criopreservação/métodos , Crioprotetores , Glucose , Inseminação Artificial/veterinária , Masculino , Sêmen/fisiologia , Preservação do Sêmen/métodos , TrometaminaRESUMO
The black-footed ferret (Mustela nigripes), which was extirpated from its native North American prairie habitat during the 1980s, is being reintroduced to the wild because of a successful captive-breeding program. To enhance propagation, the reproductive biology of this endangered species is being studied intensively. The typical life span of the black-footed ferret is approximately 7 yr. Female fecundity declines after 3 yr of age, but the influence of age on male reproduction is unknown. In this study, testis volume, seminal traits, sperm morphology, and serum testosterone were compared in 116 males from 1 to 7 yr of age living in captivity. Results demonstrated that testes volume during the peak breeding season was similar (P > 0.05) among males 1 to 5 yr of age, reduced (P < 0.05) among males 6 yr of age, and further reduced (P < 0.05) among males 7 yr of age. Motile sperm/ejaculate was similar in males 1 to 6 yr of age but diminished (P < 0.05) in those 7 yr of age. Males at 6 and 7 yr of age produced fewer (P < 0.05) structurally normal sperm than younger counterparts; however, serum testosterone concentrations were not reduced (P > 0.05) in older males. Histological comparison of testicular/epididymal tissue from 5- and 7-yr-old black-footed ferrets confirmed that the interval between these two ages may represent a transitional period to reproductive senescence. In summary, functional reproductive capacity of male black-footed ferrets exceeds that of females by at least 2 yr. Testes and seminal quality are indistinguishable among males 1 to 5 yr of age, with progressive reproductive aging occurring thereafter.
Assuntos
Envelhecimento/fisiologia , Furões/fisiologia , Sêmen/fisiologia , Espermatozoides/fisiologia , Testículo/fisiologia , Animais , Ejaculação , Epididimo/anatomia & histologia , Epididimo/fisiologia , Masculino , Motilidade dos Espermatozoides , Testículo/anatomia & histologia , Testosterona/sangueRESUMO
Acute opioid tolerance and dependence develop within hours of a single injection of morphine. We tested the hypothesis that substance P (SP) amino-terminal metabolites, originating in the periphery, affect the development of tolerance and dependence just as they modulate opioid dependence when injected intrathecally. The SP amino-terminal fragment SP(1-7) (1, 3 or 30 nmol), injected i.p. 30 min before 100 mg/kg morphine, attenuated the development of acute tolerance (5 hr) to the analgesic effect of morphine (5 mg/kg) when tested in the tail-flick assay. Injection of the D-isomer of SP(1-7), [D-Pro2, D-Phe7]SP(1-7), did not alter tolerance development but antagonized the effect of SP(1-7). Neither SP(1-7) nor [D-Pro2,D-Phe7]SP(1-7) reversed tolerance when injected 30 min before challenge with 5 mg/kg morphine. Pretreatment with SP(1-7) i.p. 30 min before 100 mg/kg morphine treatment also increased the number of withdrawal jumps induced by naloxone 3.5 hr later. In contrast, SP(1-7) given 30 min before naloxone inhibited withdrawal. [D-Pro2,D-Phe7]SP(1-7) induced an effect opposite that of SP(1-7) and antagonized the effect of SP(1-7) when coadministered. Thus, SP amino-terminal fragments have the unique characteristic of inhibiting the development of tolerance and potentiating the development but inhibiting the expression of withdrawal. These results suggest a possible mechanism by which pain-evoked release of SP may sustain opioid analgesia by attenuating the development of tolerance and inhibiting the expression of withdrawal.
Assuntos
Dependência de Morfina/tratamento farmacológico , Fragmentos de Peptídeos/uso terapêutico , Substância P/uso terapêutico , Síndrome de Abstinência a Substâncias/prevenção & controle , Animais , Relação Dose-Resposta a Droga , Tolerância a Medicamentos , Antagonistas de Aminoácidos Excitatórios/farmacologia , Injeções Intraperitoneais , Masculino , Camundongos , Naloxona/farmacologia , Fragmentos de Peptídeos/administração & dosagem , Estereoisomerismo , Substância P/administração & dosagemRESUMO
The present study was designed to determine whether the increase in N-methyl-D-aspartate (NMDA) activity in the mouse spinal cord, unmasked by naloxone in morphine-pretreated mice, is mediated by sigma receptor activity. Behavioral responses to intrathecal injections of NMDA were inhibited by pretreatment (2 h) with morphine (10 mg/kg i.p.) except when NMDA was injected together with 0.1 micrograms of naloxone. This excitatory effect of morphine on NMDA-induced behaviors, unmasked in the presence of naloxone was prevented but not reversed by haloperidol, a sigma ligand and dopamine antagonist, but not by an equivalent dose of spiperone, a dopamine antagonist. Sigma activity also appeared to contribute to morphine withdrawal jumping in mice as haloperidol inhibited naloxone-induced jumping while spiperone did not. Together these data indicate that naloxone unmasks an action of morphine on NMDA and during acute withdrawal, and these effects are each brought about by mechanisms involving sigma receptor activity.
Assuntos
Morfina/farmacologia , N-Metilaspartato/metabolismo , Receptores sigma/fisiologia , Medula Espinal/metabolismo , Animais , Comportamento Animal/efeitos dos fármacos , Haloperidol/farmacologia , Injeções Espinhais , Masculino , Camundongos , Camundongos Endogâmicos , Naloxona/farmacologiaAssuntos
Comportamento Animal/efeitos dos fármacos , Morfina/farmacologia , N-Metilaspartato/farmacologia , Receptores da Fenciclidina/efeitos dos fármacos , Receptores sigma/efeitos dos fármacos , Animais , Sinergismo Farmacológico , Agonistas de Aminoácidos Excitatórios/farmacologia , Medula Espinal/efeitos dos fármacosRESUMO
The present investigation describes the antinociceptive effect of capsaicin in the acetic acid-induced abdominal stretch assay and its mediation by substance P(1-7) fragment [SP(1-7)] and nitric oxide (NO). When injected intrathecally 24 hr before testing, SP(1-7) produced a dose-related decrease in the number of abdominal stretches induced by an i.p. injection of acetic acid. The antinociceptive effect of SP(1-7) (10 nmol) persisted for 62 hr after its injection, a time course that was similar to that produced by a dose of capsaicin (2.6 nmol) that produced an effect of similar magnitude. Antinociception induced by 10 nmol of SP(1-7) was completely reversed by coadministration of 10 nmol of D-SP(1-7); the equivalent antinociception produced by capsaicin was reversed by as small a dose as 1 nmol of D-SP(1-7). The guanylate cyclase inhibitor, methylene blue, at a dose of 10 nmol, prevented both SP(1-7)- and capsaicin-induced antinociception. Capsaicin-induced, but not SP(1-7)-induced, antinociception was prevented by Nw-nitro-L-arginine methyl ester, an NO synthase inhibitor. This inhibition of capsaicin was reversed by coadministration of 120 nmol of L-arginine. Reduced hemoglobin did not prevent capsaicin-induced antinociception. These findings suggest NO is produced and acts within capsaicin-sensitive primary afferent fibers in the dorsal spinal cord to mobilize substance P, resulting in N-terminal induced-antinociception.
Assuntos
Analgesia , Capsaicina/farmacologia , Óxido Nítrico/fisiologia , Fragmentos de Peptídeos/farmacologia , Substância P/farmacologia , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Relação Dose-Resposta a Droga , Masculino , Azul de Metileno/farmacologia , Camundongos , NG-Nitroarginina Metil ÉsterRESUMO
N-Methyl-D-aspartate (NMDA) receptors have been proposed to play a role in opioid tolerance and dependence. The present study was designed to determine whether the increased NMDA activity in the spinal cord, unmasked by naloxone in morphine-pretreated mice, reflects activity leading to opioid withdrawal. Behavioral responses to intrathecal injections of NMDA were inhibited by pretreatment (2 h) with morphine (10 mg/kg i.p.), but enhanced following morphine when naloxone was injected together with NMDA. Although injected at doses that inhibited NMDA activity, the excitatory effects of morphine on NMDA-induced behaviors were prevented by dizocilpine (MK-801), a phencyclidine (PCP) ligand, but not by 3-((+/-)-2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid (CPP), a competitive NMDA antagonist. MK-801 also inhibited naloxone-induced withdrawal jumping, however, just as CPP failed to affect morphine-induced changes in MMDA-induced behaviors, CPP also failed to inhibit withdrawal jumping. Together these data indicated that withdrawal from acute opioid dependence correlates with, but is not mediated by enhanced NMDA activity.
Assuntos
Morfina/farmacologia , N-Metilaspartato/farmacologia , Medula Espinal/fisiologia , Análise de Variância , Animais , Maleato de Dizocilpina/farmacologia , Injeções Espinhais , Masculino , Camundongos , N-Metilaspartato/administração & dosagem , N-Metilaspartato/antagonistas & inibidores , Naloxona/administração & dosagem , Naloxona/farmacologia , Piperazinas/farmacologia , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Valores de Referência , Medula Espinal/efeitos dos fármacos , Comportamento Estereotipado/efeitos dos fármacos , Síndrome de Abstinência a Substâncias/fisiopatologiaRESUMO
Substance P has been previously shown to inhibit the intensity of the morphine abstinence syndrome in mice. In view of the rapid degradation of substance P after its release from nerve terminals, we hypothesized that this inhibition is mediated by the N-terminus of substance P and its metabolites rather than via the C-terminus interacting with neurokinin receptors. Intrathecal injection of substance P-(1-7) (1 nmol) 30 min prior to naloxone challenge, in mice that had received 25 micrograms of morphine sulfate intrathecally once daily for three days, caused a dose-related attenuation of withdrawal jumping. In contrast, administration of the substance P-(1-7) antagonist, [D-Pro2,D-Phe7]substance P-(1-7), 15 min prior to naloxone increased withdrawal jumping behaviors. Equimolar doses of morphine and naloxone at 30 min had no effect. From these data, it appears that substance P N-terminal metabolites modulate withdrawal behaviors in morphine-dependent mice.