Maurizio Paro

Power spectrum analysis of cardiovascular variability monitored by telemetry in conscious unrestrained rats

Beat-to-beat variability of arterial pressure and heart period (R-R) was studied in eight conscious freely-moving adult male rats in which telemetric recordings of arterial pressure, ECG and respiratory movements were obtained under unrestrained and unstressed conditions. The beat-to-beat time series of these signals (systolic arterial pressure, diastolic arterial pressure and R-R) were analyzed, in the frequency domain, using autoregressive spectral analysis in order to detect and quantify the rhythmic components. In basal conditions, the systolic arterial pressure variability spectrum was characterized by three major spectral components which had central frequencies respectively of 0.08 +/- 0.03 Hz (very low frequency), 0.43 +/- 0.02 Hz (low frequency) and 1.36 +/- 0.19 Hz (high frequency). Similar rhythmic components were found in R-R signal variability. The very low frequency component included a higher percentage of total power in R-R variability spectrum (75.3%) than in systolic arterial pressure variability spectrum (58.4%). The low frequency component was more pronounced in both systolic and diastolic arterial pressure variability spectra. The high frequency component of R-R, systolic and diastolic arterial pressure was synchronous with respiration. Cross-spectral analysis revealed a high statistical coherence between R-R and arterial pressure variabilities in all the three frequency bands. An alpha-adrenergic blocker (phentolamine) specifically abolished the low frequency components of systolic and diastolic arterial pressure variability spectra, thus suggesting that low frequency is a marker of sympathetic modulation of vasomotor activity. The low frequency component of R-R variability spectrum was also markedly blunted. We suggest that cardiovascular variability signals, (R-R, systolic and diastolic arterial pressure) are composed almost of two main rhythms linked to respiration and vasomotor activity. These rhythms can be quantified in conscious unrestrained rats by using telemetry and spectral analysis. This approach seems to offer a new powerful tool for pharmacological studies in conscious small animals.

Cystometric changes in alloxan diabetic rats: evidence for functional and structural correlates of diabetic autonomic neuropathy

Autonomic neuropathy and urinary bladder function were compared in Sprague-Dawley rats with alloxan-diabetes of 3 months duration, rats fed sucrose for 8 weeks, and rats examined 8 weeks after pelvic nerve surgical axotomy; normal age-matched rats were used as controls. All experimental interventions induced bladder hypertrophy with increased bladder weight. In diabetic and sucrose-fed animals, water intake and urinary output increased. Cystometric recordings of normal rats in vivo showed rhythmic contractions (1.25 +/- 0.25 contr/min) with threshold volume for micturition reflex at 0.51 +/- 0.04 ml. In diabetic rats, bladder contractions were irregular and of lower frequency (0.60 +/- 0.04 contr/min), while threshold volume was significantly higher (1.00 +/- 0.11 ml). Bladder contractions were normal in sucrose-fed animals, though threshold volume was markedly augmented (1.27 +/- 0.19 ml). Pelvic nerve surgical ablation abolished micturition reflex. In bladder strips excised post-mortem, contractile response to field stimulation was reduced in diabetic rats compared to control and sucrose-fed animals. Morphological examination of pelvic and hypogastric nerves revealed abnormalities characteristic of diabetic neuropathy only in diabetic rats. These data suggest that in alloxan-induced diabetes the decrease in the rate of bladder contraction is the result of autonomic neuropathy; while bladder hypertrophy in sucrose-fed rats appears to be an organ adaptation to hyperdiuresis.

Autonomic Neuropathy in BB Rats and Alterations in Bladder Function

In vivo urinary bladder function was examined in BB rats after 4 and 6 mo of diabetes, and the data were correlated with morphometric changes in the pelvic and hypogastric nerves, which constitute the micturition reflex arc. After controlled bladder distension, diabetic animals revealed irregular bladder contractions at frequencies that were reduced to 33% of normal values and with significantly increased amplitudes. The abnormal micturition in diabetic animals was elicited at moderately elevated threshold volumes. These functional abnormalities of the diabetic bladder were associated with a progressive axonopathy of afferent myelinated sensory fibers and later-occurring axonal atrophy of unmyelinated efferent preganglionic fibers. These data suggest that diabetic urinary bladder dysfunction is initiated by a visceral sensory neuropathy involving the afferent limb of the micturition reflex arc.

Urinary Bladder Dysfunction in the BB/W Diabetic Rat: Effect of Ganglioside Treatment on Functional and Structural Alterations

Urinary bladder dysfunction in the diabetic BB/W rat is characterized by infrequent irregular contractions of high amplitude. Initially these occur in the absence of detectable neuroanatomical lesions of sensory afferent and parasympathetic fibers of the pelvic nerve, which constitute the micturition reflex arc. Structural lesions consisting of progressive axonal atrophy of myelinated and unmyelinated fibers become detectable only after 4 months of diabetes. In the current study we evaluated the effect of ganglioside treatment (10 mg./kg. body weight) for one month. This drug regimen was initiated at 4 months of diabetes, when functional bladder abnormalities were well established, whereas structural lesions were yet to appear. Animals examined 1 or 3 months after termination of the one-month treatment protocol showed sustained normalization of the characteristic functional abnormalities, accompanied by prevention of the neuroanatomical lesions of sensory afferent and parasympathetic efferent myelinated fibers in the pelvic nerve. These data suggest that ganglioside treatment may be beneficial in delaying the progression of diabetic autonomic neuropathy in this experimental animal model.

Experimental diabetes in the rat: alterations in the vesical function

Vesical function has been studied in vivo in alloxan-diabetic rats. Rhythmic contractions evoked as a micturition reflex by a controlled distension of the bladder were recorded in urethane-anesthetized animals. These rhythmic contractions are essentially induced by the parasympathetic and inhibited by the sympathetic outflow. The study has been performed on male Sprague-Dawley rats. They were treated at the age of two months with vehicle (control group) or alloxan (100 mg/kg s.c.). Such treatment caused hyperglycemia (greater than 250 mg/dl) and glycosuria in most of the animals. In control animals, analyzed between 5 and 8 months of age, the contractions evoked by vesical distension appeared at a threshold volume between 0.5 and 1 ml, and they had a regular rate. In a first series of experiments, diabetic animals 3 months after treatment showed the evoked response at an average threshold volume of about 1.7 ml. while in animals with 6 months of experimental diabetes the average threshold volume was about 3.2 ml. The rate of contractions in diabetic animals became more and more irregular as the duration of diabetes increased. These marked functional alterations were associated with the evidence of bladder enlargement. In a second series of experiments, bovine brain gangliosides (10 mg/kg i.p./day) or saline were given to diabetic rats between the 30th and the 90th day after alloxan. Ganglioside administration significantly reduced bladder enlargement and the threshold volume for micturition reflex and improved the rate of bladder contractions. It is suggested that alterations of the autonomic nervous system can be expressed as functional vesical dysfunction in experimental diabetes.