Reflex Modification Audiometry Research Articles

A BK channel-targeted peptide induces age-dependent improvement in behavioral and neural sound representation

Recent evidence suggests that modulation of the large-conductance, calcium-activated potassium (BK) channel regulates auditory processing in the brain. Because ion channel expression often changes during aging, this could be a factor in age-related hearing loss. The current study explored how the novel BK channel modulator LS3 shapes central auditory processing in young and old adult mice. In vivo extracellular recordings in the auditory midbrain demonstrated that LS3 differentially modulates neural processing along the tonotopic axis. Though sound-evoked activity was reduced in the mid and ventral tonotopic regions, LS3 enhanced excitatory drive and sound-evoked responses for some neurons in the dorsal, low-frequency region. Behavioral assessment using acoustic reflex modification audiometry indicated improved tone salience following systemic LS3 administration. Moderation of these responses with aging correlated with an age-related decline in BK channel expression. These findings suggest that targeting the BK channel enhances responsivity to tonal sounds, providing the potential to improve hearing acuity and treat hearing loss.

Reflex Modification Audiometry Reveals Dual Roles for Olivocochlear Neurotransmission

Approximately 15% of American adults report some degree of difficulty hearing in a noisy environment or have auditory filtering difficulties. There are objective clinical tests of auditory filtering, yet few tests exist for mouse models that do not rely on extensive training. We have used reflex modification audiometry (RMA) and developed exclusion criteria for the mouse model. This RMA based test makes use of the acoustic startle response (ASR) and the ability of prepulses to inhibit the ASR [i.e., prepulse inhibition (PPI)] to assess the mouse's ability to detect prepulse signals presented in quiet or embedded in masking noise. We have studied PPI behavior across four inbred mouse strains with normal cochlear function and developed pre-testing exclusion criteria and test/retest reliability measures. Moreover, because both the medial (MOC) and the lateral (LOC) olivocochlear efferent feedback systems have been proposed to improve auditory behavior performance, especially in noisy backgrounds, we have examined PPI abilities in mice (with their littermate controls) either lacking the MOC receptor subunit α9 nicotinic acetylcholine receptor [α9 nAChR (–/–)] or expressing an overactive receptor [Ld'T mutation in α9 nAChR KI], or lacking an LOC efferent neuropeptide, alpha calcitonin gene-related peptide [αCGRP (–/–)] only in the CNS. Because CGRP receptor formation has been shown to mature from juvenile to adult ages, we also studied if this maturation would be reflected in PPI behavioral responses in juvenile and adult (+/+) controls and in adult αCGRP (–/–) animals. We show that 50% PPI response thresholds (sound level with 50% correct responses) in quiet are decreased in the (–/–) α9 nAChR animals, and 50% PPI responses are increased for mice with an overactive receptor (α9 nAChR KI) and are increased in adult mice lacking αCGRP (–/–). However, in background noise, only mice lacking αCGRP exhibited increased 50% PPI response thresholds, as there were no significant differences between α9 nAChR adult mouse lines and their littermate controls. These findings suggest that MOC and LOC olivocochlear neurotransmission work in tandem to improve behavioral responses to sound. These experiments further pave the way for rapid behavioral hearing assessments in other mouse models.

Kcna1Gene Deletion Lowers the Behavioral Sensitivity of Mice to Small Changes in Sound Location and Increases Asynchronous Brainstem Auditory Evoked Potentials But Does Not Affect Hearing Thresholds

Sound localization along the azimuth depends on the sensitivity of binaural nuclei in the auditory brainstem to small differences in interaural level and timing occurring within a submillisecond epoch and on monaural pathways that transmit level and timing cues with high temporal fidelity to insure their coincident arrival at the binaural targets. The soma and axons of these brainstem neurons are heavily invested with ion channels containing the low-threshold potassium channel subunit Kv1.1, which previous in vitro and in vivo studies suggest are important for regulating their high input-output correspondence and temporal synchrony. We compared awake Kcna1-null mutant (Kcna1-/-) mice lacking Kv1.1 with Kcna1+/+ mice to determine whether Kv1.1 activity contributes to sound localization and examined anesthetized mice for absolute hearing thresholds for suprathreshold differences that may be revealed in the waveforms of auditory brainstem response potentials. The awake -/- mice tested with reflex modification audiometry had reduced sensitivity to an abrupt change in the location of a broad band noise compared to +/+ mice, while anesthetized -/- mice had normal absolute thresholds for tone pips but a high level of stimulus-evoked but asynchronous background activity. Evoked potential waveforms had progressively earlier peaks and troughs in -/- mice, but the amplitude excursions between adjacent features were identical in the two groups. Their greater excitability and asynchrony in suprathreshold evoked potentials coupled with their normal thresholds suggests that a disruption in central neural processing in -/- mice and not peripheral hearing loss is responsible for their poor sound localization.

Ototoxicity

Ototoxicity

The relative detectability for mice of gaps having different ramp durations at their onset and offset boundaries.

The effect on gap detectability of varying noise fall time (FT) and rise time (RT) of the gap boundary ramps was examined in mice using reflex modification audiometry, measuring inhibition of acoustic startle reflexes by variously shaped gaps just preceding reflex expression. In experiment 1 (n = 12) inhibition increased up to near-asymptotic values with longer FT (0, 1, 2, 3, 5, or 10 ms) and QT (quiet time, 0 to 13 ms), with a 2:1 trade-off between FT and QT. In experiment 2 (n = 24) inhibition increased for any RT above 0 ms (2, 3, 5, or 7 ms) if QT= 1 ms, but diminished with increased RT when QT = 3 or 8 ms. Enhanced detectability for subthreshold gaps by longer ramps results from their extending the apparent gap duration. The negative effect of increased RT for threshold gaps suggests the importance for gap detection of the stronger neural responses to sharp edges at the end of the gap shown previously in the mouse inferior colliculus. These effects are specific to gaps: inhibition for fixed (70-dB SPL) or varied level pulses (30 to 60 dB) was unaffected by varying the ramped edges (experiments 3 and 4, n = 9).

Flash-, somatosensory-, and peripheral nerve-evoked potentials in rats perinatally exposed to Aroclor 1254

Pregnant Long–Evans rats were exposed to 0, 1 or 6 mg/kg/day of Aroclor 1254 (A1254; Lot no. 124-191), a commercial mixture of polychlorinated biphenyls (PCBs), from gestation day (GD) 6 through postnatal day (PND) 21. At 128–140 days of age, male and female offspring were tested for visual-, somatosensory- and peripheral nerve-evoked potentials. The evoked responses increased in amplitude with larger stimulus intensities, and gender differences were detected for some endpoints. In contrast, developmental exposure to A1254 failed to significantly affect the electrophysiological measures. A subset of the animals were tested for low-frequency hearing dysfunction using reflex modification audiometry (RMA). An elevated threshold for a 1-kHz tone was observed, replicating previous findings of A1254-induced auditory deficits [Hear. Res. 144 (2000) 196; Toxicol. Sci. 45(1) (1998) 94; Toxicol. Appl. Pharmacol. 135(1) (1995) 77.]. These findings indicate no statistically significant changes in visual-, somatosensory- or peripheral nerve-evoked potentials following developmental exposure to doses of A1254 that produce behavioral hearing deficits. However, subtle changes in the function of the visual or somatosensory systems cannot be disproved.

Scopolamine reduces sensitivity to auditory gaps in the rat, suggesting a cholinergic contribution to temporal acuity

Prior research [Caine et al., 1981] suggested that scopolamine, a central cholinergic antagonist, may increase gap thresholds in young human listeners. If confirmed, an effect of scopolamine on gap detection might help to explain why both aged humans and aged laboratory animals have less sensitive temporal acuity on gap detection tests, as they may be presumed to have less effective cholinergic mechanisms. Here we measured the effect of scopolamine on gap detection in rats ( n=8) using reflex modification audiometry, which depends on the fact that brief gaps in noise presented immediately prior to a loud noise inhibit the acoustic startle reflex. Scopolamine increased the gap threshold and reduced reflex inhibition produced by gaps that were presented at and beyond about 40 ms prior to the startle reflex, but not at shorter lead times. A peripheral antagonist had no effect at long lead times. These data indicate that central cholinergic mechanisms are involved in relatively high level perceptual processing of gaps. This conclusion is consistent with the hypothesis that temporal acuity may be compromised in the aged listener because of deficits in the efficacy of these central mechanisms.

Hearing loss following exposure during development to polychlorinated biphenyls: A cochlear site of action

Maternal exposure to polyhalogenated hydrocarbons results in early postnatal hypothyroxenemia and a low-frequency hearing loss in adult offspring (Goldey et al., 1995a. Toxicol. Appl. Pharmacol. 135, 67–76; Herr et al., 1996. Fundam. Appl. Toxicol. 33, 120–128). The purpose of the present work was to determine whether the site-of-action of this auditory impairment was within the cochlea. Primiparous Long–Evans rats were given daily oral doses of corn oil (control) or 8 mg/kg of the commercial PCB mixture Aroclor 1254 (A1254) from gestation day (GD) 6 through postnatal day (PND) 21. Auditory thresholds for 1-, 4-, 16-, and 40-kHz tones were assessed using reflex modification audiometry in young adult offspring on postnatal days (PND) 92–110. Approximately 6 weeks after auditory assessments, a subset of animals ( n=4 per group) were killed for histological assessment of the cochlea. Surface preparations of the organ of Corti were prepared from one cochlea per animal and modiolar sections were prepared from the opposite cochlea. Consistent with previous findings, auditory thresholds for 1-kHz tones were elevated by approximately 25 dB in the A1254-exposed animals. Thresholds for all higher frequencies were not different compared to controls. Surface preparations of the organ of Corti revealed a mild to moderate loss of outer hair cells in the upper-middle and apical turns. Inner hair cells were not affected. Modiolar sections failed to reveal alterations in any other cochlear structures. There was also no apparent loss of ganglion cells. These data clearly link the loss of low-frequency hearing caused by exposure during development to A1254 to a loss of outer hair cells in the organ of Corti. The mechanism that underlies this developmental ototoxicity remains to be determined. These data provide the first evidence of a structural deficit in the nervous system of adult animals exposed to PCBs during development.

The ototoxic effects of ethyl benzene in rats

Exposure to organic solvents has been shown to be ototoxic in animals and there is evidence that these solvents can induce hearing loss in humans. In this study, the effects of inhalation of the possibly ototoxic solvent ethyl benzene on the cochlear function and morphology were evaluated using three complementary techniques: (1) reflex modification audiometry (RMA), (2) electrocochleography and (3) histological examination of the cochleas. Rats were exposed to either ethyl benzene (800 ppm, 8 h/day for 5 days) or to control conditions. The RMA threshold increased significantly by about 25 dB, 1 and 4 weeks after the exposure, irrespective of the stimulus frequency tested (4–24 kHz). Electrocochleography was performed between 8 and 11 weeks after exposure to the organic solvent. The threshold for the compound action potential increased significantly by 10–30 dB at all frequencies tested (1–24 kHz). Histological examination of the cochlea showed outer hair cell (OHC) loss, especially in the upper basal and lower middle turns (corresponding to the mid-frequency region) to an extent of 65%. We conclude that exposure to 800 ppm ethyl benzene for 8 h/day during 5 days induces hearing loss in rats due to OHC loss.

Changes in temporal acuity with age and with hearing impairment in the mouse: a study of the acoustic startle reflex and its inhibition by brief decrements in noise level.

Temporal acuity for brief gaps in noise was studied in mice of different ages (1-36 months) from strains with differing susceptibility to age-related hearing loss, using reflex modification audiometry. Prepulse inhibition of the acoustic startle reflex (ASR) increased with gap depth (GD: 10-40 dB in 70 dB SPL noise) and lead time (LT: 1-15 ms). The increase in inhibition with LT followed an exponential function in which the two parameters, asymptotic inhibition (AINH) and the time constant (tau), were both affected by GD. AINH rapidly declined from 1 to 6 and then to 18 months of age in C57BL/6J mice with progressively severe hearing loss, but first increased with maturation and then gradually declined beyond 6-12 months of age in CBA/CaJ and CBA x C57BL Fl-hybrid mice, which show no apparent change in sensory function at these ages. In contrast, tau was unaffected by hearing loss or by age, this suggesting that age-related changes in this form of temporal acuity occur because of a reduction in the efficiency with which gaps are centrally processed, not from any reduced ability to follow their rapid shift in noise level.

The ototoxicity of trichloroethylene: extrapolation and relevance of high-concentration, short-duration animal exposure data.

Inhalation exposure to high concentrations of 1,1, 2-trichloroethylene (TCE) has been shown to damage hearing in the mid-frequency range in the rat. The present study directly evaluated the adequacy of high-concentration, short-term exposures to TCE for predicting the neurotoxicity produced by longer duration exposures. Adult male Long-Evans rats (n = 10-12 per group) were exposed to TCE via inhalation (whole body) in 1-m3 stainless steel flow-through chambers for 6 hr/day, 5 days/week. The following exposures were used: 1 day (4000-8000 ppm), 1 week (1000-4000 ppm), 4 weeks (800-3200 ppm), and 13 weeks (800-3200 ppm). Air-only exposed animals served as controls. Auditory thresholds were determined for a 16-kHz tone 3-5 weeks after exposure using reflex modification audiometry. Results replicated previous findings of a hearing loss at 16 kHz for all exposure durations. The dB15 concentrations (concentration that increases thresholds by 15 dB) for 16-kHz thresholds were 6218, 2992, 2592, and 2160 ppm for the 1-day, 1-week, 4-week and 13-week exposures, respectively. These data demonstrate that the ototoxicity of TCE was less than that predicted by a strict concentration x time relationship. These data also demonstrate that simple models of extrapolation (i.e., C x t = k, Haber's Law) overestimate the potency of TCE when extrapolating from short-duration to longer-duration exposures. Furthermore, these data suggest that, relative to ambient or occupational exposures, the ototoxicity of TCE in the rat is a high-concentration effect.

The Ototoxicity of Trichloroethylene: Extrapolation and Relevance of High-Concentration, Short-Duration Animal Exposure Data

Inhalation exposure to high concentrations of 1,1,2-trichloroethylene (TCE) has been shown to damage hearing in the mid-frequency range in the rat. The present study directly evaluated the adequacy of high-concentration, short-term exposures to TCE for predicting the neurotoxicity produced by longer duration exposures. Adult male Long-Evans rats (n = 10–12 per group) were exposed to TCE via inhalation (whole body) in 1-m3 stainless steel flow-through chambers for 6 hr/day, 5 days/week. The following exposures were used: 1 day (4000–8000 ppm), 1 week (1000–4000 ppm), 4 weeks (800–3200 ppm), and 13 weeks (800–3200 ppm). Air-only exposed animals served as controls. Auditory thresholds were determined for a 16-kHz tone 3–5 weeks after exposure using reflex modification audiometry. Results replicated previous findings of a hearing loss at 16 kHz for all exposure durations. The dBl5 concentrations (concentration that increases thresholds by 15 dB) for 16-kHz thresholds were 6218, 2992, 2592, and 2160 ppm for the 1-day, 1-week, 4-week and 13-week exposures, respectively. These data demonstrate that the ototoxicity of TCE was less than that predicted by a strict concentration × time relationship. These data also demonstrate that simple models of extrapolation (i.e., C × t = k, Haber's Law) overestimate the potency of TCE when extrapolating from short-duration to longer-duration exposures. Furthermore, these data suggest that, relative to ambient or occupational exposures, the ototoxicity of TCE in the rat is a high-concentration effect.

Assessment of Offspring Development and Behavior Following Gestational Exposure to Inhaled Methanol in the Rat

Assessment of Offspring Development and Behavior Following Gestational Exposure to Inhaled Methanol in the Rat. Stanton, M. E., Crofton, K. M., Gray, L. E., Gordon, C. J., Boyes, W. K., Mole, M. L., Peele, D. B, and Bushnell, P. J. (1995). Fundam. Appl. Toxicol. 28, 100-110. The prospect of widespread human exposure associated with its use as an alternative fuel has sparked concern about the toxic potential of inhaled methanol (MeOH). Previous studies have revealed congenital malformations in rats following inhaled MeOH (Nelson et al. (1985). Fundam. Appl. Toxicol. 5, 727-736) but these studies did not include postnatal behavioral assessment. In the present study, pregnant Long-Evans rats were placed in exposure chambers containing 15,000 ppm MeOH or air for 7 hr/day on Gestational Days (GD) 7-19. The total alveolar dose of methanol was estimated at about 6.1 g/kg/day, for a total dose of about 42.7 g/kg for the entire study. Maternal body weights were recorded daily and blood methanol concentrations were determined at the end of exposure on GD 7, 10, 14, and 18. Following birth (Postnatal Day 0 [PND 0]), a number of tests were performed at various points in development, including: offspring mortality and body wt (PND 1, 3), motor activity (PND 13-21, 30, 60), olfactory learning (PND 18), behavioral thermoregulation (PND 20-21), T-maze learning (PND 23-24), acoustic startle response (PND 24, 60), reflex modification audiometry (PND 60), pubertal landmarks (PND 31-56), passive avoidance (PND 72), and visual-evoked potentials (PND 160). Maternal blood MeOH levels, measured from samples taken within 15 min after removal from the exposure chamber, declined from about 3.8 mg/ml on the first day of exposure to 3.1 mg/ml on the 12th day of exposure. MeOH transiently reduced maternal body wt (4-7%) on GD 8-10, and offspring BW (5%) on PND 1. No. other test revealed significant effects of MeOH. Prenatal exposure to high levels of inhaled MeOH appears to have little effect on this broad battery of tests beyond PND 1 in the rat.

Effects of Developmental Hypothyroidism on Auditory and Motor Function in the Rat

Deafness is a common result of severe hypothyroidism during development in humans and laboratory animals; however, little is known regarding the sensitivity of the auditory system to more moderate changes in thyroid hormone homeostasis. The current investigation compared the relative sensitivity of auditory function, motor function, and growth to the effects of moderate to severe perinatal hypothyroidism in the rat. Rats received propylthiouracil (PTU) in drinking water at concentrations of 0, 1, 5, and 25 ppm from Gestation Day 18 until postnatal day (PND) 21, and the effects on their offspring were evaluated. At 1 ppm, PTU did not affect any of the measured endpoints. Serum thyroxin concentrations were sharply reduced in the 5 and 25 ppm PTU groups at all ages sampled (PND 1, 7, 14, and 21). Marked reductions in serum triiodothyronine (T3) concentrations were also detected for all ages ≥7 at 25 ppm PTU, whereas no effects of 5 ppm PTU on serum T3 were apparent until PND 21. Compared to the controls, pups exposed to the highest dose of PTU demonstrated a delay in eye opening, reduced body weights, decreased and/or delayed preweaning motor activity, and persistent, postweaning hyperactivity. Only slight and transient effects on eye opening and ontogeny of motor activity were seen at the intermediate dose of PTU (5 ppm). Reflex modification audiometry revealed that, compared to controls, adult offspring from the 5 and 25 ppm treatment groups showed dose-dependent auditory threshold deficits (35 to >50 dB) at all frequencies tested (1, 4, 16, 32, and 40 kHz). Such dose-dependent effects indicate that the developing auditory system may be sensitive to mild hypothyroidism, suggesting the possible need for routine audiometric screening for infants and children at risk for iodine deficiency, myxedema, and/or exposure to thyrotoxic environmental agents.

Solvent-induced ototoxicity in rats: An atypical selective mid-frequency hearing deficit

Most previous reports of ototoxicity following exposure to several volatile organic solvents have restricted testing to the low-and mid-frequencies (2–20 kHz) of the hearing range in the rat (0.25–80 kHz). We report here that inhalation exposure to styrenc, mixed xylene, toluene, and 1,1,2-trichloroethylene resulted in hearing dysfunction only in the mid-frequency range and spared function at lower and higher frequencies. Adult male Long Evans rats were exposed via inhalation (whole body) in flow-through chambers. The following exposures were used: styrene, 1600 ppm; 1,1,2-trichloroethylene, 3500 ppm; toluene, 2500 ppm; mixed xylenes, 1800 ppm ( N = 7−8 per group, 8 h/day for 5 days), and n-butanol, 4000 ppm ( N = 10/group, 6 h/day for 5 days). Testing of auditory function was conducted 5 to 8 weeks after exposure using reflex modification audiometry (RMA). RMA thresholds were determined for frequencies from 0.5 to 40 kHz. Results indicated increased RMA thresholds for the mid-frequency tones (e.g., 8 and 16 kHz), but not higher or lower tones, for all solvents except n-butanol. Toluene and xylene also increased thresholds at 24 kHz. These data indicate that for those solvents reported thus far to cause hearing loss, the deficit is restricted to mid-frequencies in rats.

Mid-frequency hearing loss in rats following inhalation exposure to trichloroethylene: Evidence from reflex modification audiometry

The present experiments were undertaken to characterize the hearing loss associated with 1,1,2-trichloroethylene (TCE) exposure. Adult male Long-Evans (LE) rats were exposed to TCE via inhalation (whole body) for 6 h/day for 5 days. The concentration-effect function (0–4000 ppm) was determined 3 weeks post-exposure. Animals were tested for auditory thresholds to 4, 8, 16, 24, 32, and 40-kHz tones using reflex modification audiometry. In a separate experiment, the time course of effects was determined by monitoring 16-kHz thresholds prior to, 1 h following each of the 5 exposure days, and 5 days, 1, 2, 4, 8, and 12 weeks post-exposure. At 14 weeks, these same animals were tested for thresholds to 0.5, 1, 2, 4, 8, 16, 24, 32, and 40-kHz tones. Results indicate elevated thresholds (hearing loss) for the 4000 ppm group at 8 and 16 kHz of approximately 18 and 30 dB, respectively. Time-course data demonstrated a rapid onset, a 20-dB loss at 16 kHz after the fifth exposure day, and a 40-dB loss by 2 weeks that persisted up to 14 weeks post-exposure. These data demonstrate an atypical and persistent, mid-frequency hearing loss in rats following inhalation exposure to TCE.

The sensitivity to 3,3′-iminodipropionitrile differs for high-and midfrequency hearing loss in the developing rat

3,3′-Iminodipropionitrile (IDPN) has been demonstrated to produce a loss of hearing following both neonatal and adult exposures. Adult exposure induces a full spectrum hearing loss, whereas early postnatal exposure produces a high-frequency loss only. The purpose of this work was to delineate the period of development during which the rat becomes sensitive to the full ototoxic effects of IDPN. Primiparous Long Evans rats or their offspring were exposed to either saline or 300 mg/kg IDPN for three consecutive days. Ages of exposure were as follows: gestational days 15–17 or postnatal days (PND) 1–3, 5–7, 15–17, 20–22, 25–27, 30–32, 40–42, or 70–72. All animals were tested as adults for auditory thresholds to 5- and 40-kHz tones using reflex modification audiometry. Results demonstrate that adult-like susceptibility to IDPN was not reached until approximately PND 30–32. Early exposures (PND 5–22) to IDPN will induce a highfrequency selective hearing loss, sparing the lower frequency. Prenatal or early neonatal (PND 1–3) IDPN exposure resulted in a high degree of mortality ( $ ̆ 70%). The long period of time between the susceptible period for the high frequency (PND 5–7) and the lower frequency (PND 30–32) does not correspond to the basal to apical ontogenic profile of any one physiological or anatomical process. These data suggest either a unique site of action for IDPN in the cochlea or the possibility of two different mechanisms, one operating at early postnatal ages and one at later ages.

Sensory threshold estimation from a continuously graded response produced by reflex modification audiometry.

This article describes the use of reflex modification to determine sensory detection thresholds. The method is based upon the finding that low-intensity sensory stimuli presented shortly before a reflex eliciting stimulus are able to modify the amplitude of the reflex. The extent of such modification is related to the intensity of the initial low-intensity stimulus. In contrast to earlier reported procedures for threshold estimation, the method described in this article consists of fitting a smooth function to the relationship between startle response amplitude and the intensity of the inhibiting stimulus. The method entails fitting a cubic spline function to the medians of the square-root reflex amplitude at each prestimulus intensity. The resulting audiometric curves closely approximate audiometric data obtained from traditional operant methods both in sensitivity and shape. Parametric data are also presented that allow for optimizing stimulus presentation so as to obtain reliable thresholds using a minimal number of test trials. The procedures developed in this article may prove useful in other situations involving the estimation of a threshold effect from a continuously graded response.

COMPARISON OF REFLEX MODIFICATION AUDIOMETRY AND BRAINSTEM EVOKED RESPONSE AUDIOMETRY IN HIGH–RISK PREMATURE INFANTS

Reflex modification audiometry (RMA) is an objective technique for assessing hearing acuity by determining whether the reflexive eyeblink, elicited by a tap to the glabella, is augmented by an acoustic stimulus presented simultaneously with the glabellar tap. Sixteen infants who were at risk for hearing impairment were tested with both RMA and brainstem evoked response audiometry (ABR). These infants had either intraventricular hemorrhage, meningitis, prolonged amingoglycoside therapy and/or were small for gestational age. The RMA procedure consisted of the elicitation of an eyeblink by a miniature solenoid with a teflon striker which could deliver a mild controlled tap to the glabella. The tap was presented either alone or simultaneously with a tone at 90 dB SPL. A photoreflective densitometer attached to a TDH39 earphone provided an objective measure of eyeblink amplitude. The ABR was obtained through standard procedures using clicks at 60 dB HL. Eleven infants passed the brainstem procedure (Group 1: mean postconceptional age 38 wks) while 5 infants failed (Group 2: mean postconceptional age 34 wks). A comparison of the ABR with the reflex augmentation testing procedure showed a significant difference (p<0.05) between those infants who passed the ABR and showed augmentation (10/11 showed augmentation, 4/11 at p<0.05) and those who failed the ABR and showed no difference in their response to tap alone and tap with tone. These data suggest that reflex modification audiometry, which is inexpensive and requires little training and less time to administer and interpret, may be a promising tool for screening infants at risk for hearing loss.