Margarita Martínez-Gómez

Assessing pain threshold in the rat: Changes with estrus and time of day

Pain threshold was determined in female rats using the tail flick test. Latency to respond depended on the locus of the tail heated, with the most distal sites resulting in the shortest response times (Experiment 1). Tail flick latency also varied according to the time of day, with shorter response times recorded around the middle of the dark phase than near its beginning or after its end. This was the case for intact, for ovariectomized, and for ovariectomized, estradiol-treated rats (Experiment 2). However, response times also varied across the estrous cycle, with significantly shorter latencies recorded during estrus and metestrus. Ovariectomy abolished these fluctuations, and whereas administration of estradiol increased response times, progesterone had little effect (Experiment 3). These results suggest that site of tail heating, time of day, and presence of ovarian hormones can influence tail flick latency independently, thus demonstrating the complexity of the mechanisms that may contribute to pain threshold even within the same paradigm.

Metabolic correlates of the circadian pattern of suckling-associated arousal in young rabbits

Abstract Young rabbits (Oryctolagus cuniculus) are only nursed for 3–5 min every 24 h. They show a circadian increase in activity in anticipation of this, which is entrained by suckling. Our aim was to determine whether serum and liver metabolites show diurnal fluctuations which could act to regulate this circadian pattern. Stomach weight, liver glycogen and serum metabolites were measured every 3 h in 7- to 8-day-old pups when normally nursed (up to 24 h after suckling) and fasted (up to 48 h after suckling). The results suggest:

Striated muscles and scent glands associated with the vaginal tract of the rabbit

Despite the importance of the rabbit (Oryctolagus cuniculus) in reproductive research, no comprehensive account of the anatomical organization of the female reproductive tract is presently available.

Visceral and postural reflexes evoked by genital stimulation in urethane-anesthetized female rats

The present study describes several muscular reflexes produced by genital stimulation, the nerves that subserve them, and the visceral and postural effects induced by these reflexes. Electrical stimulation of the iliococcygeus (ic) and pubococcygeus (pc) (striated) muscles produced movement of the vaginal orifice and wall, membranous urethra, tail and pelvis. Electrical stimulation of the psoas major (pm) or iliacus (i) (striated) muscles produced movements of the lumbar vertebrae and extension of the ipsilateral hindlimb. Sensory mechanostimulation elicited responses of these muscles as follows: stimulation of the perineal skin, clitoral sheath or distal vagina produced reflex contraction of the ic and pc muscles. Stimulation of the cervix produced reflex contraction of the pm and i muscles and also blocked the above reflex contraction of the ic and pc muscles. Both the cervical stimulation-induced blockage of the ic and pc reflex response, and the cervical stimulation-induced activation of pm and i muscles was prevented by bilateral transection of the viscerocutaneous branch of the pelvic nerve. Based on the above observations, it is proposed that stimulation of the vaginal surface of the cervix resulting from penile intromission and/or seminal plug deposition during mating behavior in the rat may reflexively active pm and i, thereby contributing to the hindleg postural rigidity and lordotic dorsiflexion that are characteristic of the normal mating posture in female rats.

Participation of pelvic nerve branches in male rat copulatory behavior

The role of the pelvic nerve branches in the mediation of copulatory behavior was investigated. The somatomotor or the viscerocutaneous branch of the pelvic nerve was bilaterally sectioned in sexually experienced male rats. Somatomotor branch surgery had no detectable effect. Viscerocutaneous branch transection altered copulatory parameters that reflect impairments in penile erection and seminal plug emission. The altered behavioral parameters approached or reached presurgical and sham values 21 days after transection, indicating that the damage to erectile and ejaculatory function was transient. It is suggested that animals with viscerocutaneous branch transection recover copulatory efficiency through a compensatory plastic mechanism, possibly involving the hypogastric nerve.

Changes in pain threshold during the reproductive cycle of the female rat.

Responsiveness to pain was determined in female rats across the whole reproductive cycle using the tail-flick test. When tested immediately after mating, pain thresholds were unaltered, whereas 10 min later animals typically demonstrated hyperalgesia (Experiment 1). They also demonstrated hyperalgesia during most of pregnancy, and had significantly lower pain thresholds than the unmated controls except for the 24 h before parturition, when a sudden increase in tail-flick latencies was recorded (Experiment 2). Pain thresholds were also significantly lower throughout the nursing period but increased significantly when dams were separated from their litters for 6 h, and returned to premating baseline values within 24 h of weaning (Experiment 3). These findings confirm and extend earlier reports that female reproductive state may significantly modify responsiveness to noxious stimuli, and it is suggested that differences between the results of this and previous studies may be at least partly explained by the relatively stress-free test procedure adopted here.

An anatomical and electrophysiological study of the genitofemoral nerve and some of its targets in the male rat.

Anatomical descriptions of the genitofemoral nerve (GFn) innervating the lower pelvic area are contradictory. Here we re‐examine its origin and innervation by its various branches of principal target organs in the male rat. Using gross dissection, electrophysiological techniques and retrograde tracing of motoneurones with horseradish peroxidase, we confirm that the GFn originates from lumbar spinal nerves 1 and 2, and that at the level of the common iliac artery it divides into a lateral femoral and a medial genital branch. In contrast to previous studies, we report that the genital and not the femoral branch innervates the abdominal–inguinal skin, and not only the genital but also the femoral branch innervates the cremaster muscle (Cm) surrounding the testes. Motoneurones innervating the Cm proper are located in the ventral nucleus of L1 and L2, and those innervating the muscular transition region of the rostral Cm are located in the ventral nucleus in L1 and the ventrolateral nucleus in L2. The GFn may contribute to male reproductive performance by transmitting cutaneous information during copulation and, via contraction of the Cm to promote ejaculation, the protective displacement of the testes into the abdominal cavity during fighting and as a sperm‐protecting thermoregulatory measure.

Pattern of sensory innervation of the perineal skin in the female rat.

Here we describe the nerves innervating the perineal skin together with their sensory fields in the adult female rat. Electrophysiological recording showed that the lumbosacral and L6-S1 trunks, in part by way of the sacral plexus, transmit sensory information from the perineal skin via four nerves: the viscerocutaneous branch of the pelvic nerve innervating the skin at the midline between the vaginal opening and anus, the sensory branch of the pudendal nerve innervating the clitoral sheath, the distal perineal branch of the pudendal nerve innervating a broad area of skin adjacent to the vaginal opening and anus, and the proximal perineal branch of the sacral plexus innervating a broad area of skin adjacent to the clitoris and vaginal opening. The sensory fields of three of these nerves overlapped to some degree: the viscerocutaneous branch of the pelvic and the distal perineal branch of the pudendal nerves at the midline skin between the vaginal opening and the anus, and the distal perineal branch of the pudendal nerve and the proximal perineal branch of the sacral plexus at the skin lateral to the vaginal opening. Such overlap might provide a safeguard helping to ensure that somatosensory input from the perineal region important for triggering reproductive and nonreproductive reflexes reaches the CNS.

Anatomical and physiological characteristics of perineal muscles in the female rabbit

Little information is available on the participation of the perineal striated muscles in female reproductive processes. Here, we describe the gross anatomy and innervation of two striated perineal muscles in the female rabbit, the bulbospongiosus (BSM) and ischiocavernosus (ISM), and analyze their reflex electromyographic (EMG) activity in response to stimulation of the perigenital skin and vaginal tract. Twenty-four mature chinchilla-breed rabbit does were used: 12 to describe the anatomy and innervation of the muscles, 9 to determine reflex EMG activity of the muscles in response to stimulation of the perigenital skin and specific levels of the vaginal tract and 3 to analyze the effect of contraction of the muscles on intravaginal pressure. Both muscles were well developed, with their fibers originating at the ischiadic arch and inserting onto the ligamentum suspensorium clitoridis. Branches of the clitoral and perineal nerves innervated the BSM and ISM, respectively. Bilateral electrical stimulation of these nerves provoked retraction of the clitoral sheath and an increase in intravaginal pressure at the level of the perineal vagina. Whereas neither muscle responded to stimulation of the perigenital skin, both were reflexively activated during mechanical stimulation of the inner walls of the perineal vagina. Prolonged cervical stimulation inhibited this reflex. Thus, in reproductive processes such as copulation and/or parturition, the contraction of these muscles may be induced during stimulation of the perineal vagina.

The Role of Pubococcygeus Muscle in Urinary Continence in the Male Rat

The role of the male rat pubococcygeus muscle (Pcm) in the micturition reflex was analyzed. Anatomical features of Pcm, electrical stimulation of its nerve, electrical recording and stimulation of the muscle and cystometrograms were carried out. Results showed that Pcm has fibers attached to the ventrolateral part of the external urethral sphincter, and that its activity contributes to hold the tail in the midline. Pcm shows activity during fluid expulsion in cystometrograms and spontaneous micturitions. This activity produced reflex inhibition of detrusor contraction and was not the cause of intravesical high frequency oscillations. Thus, it is proposed that Pcm activity produces the discharge of its afferents which in turn activates a spinal reflex to promote continence.