<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2024-06-17</periodOfReport>

    <issuer>
        <issuerCik>0001617867</issuerCik>
        <issuerName>Autonomix Medical, Inc.</issuerName>
        <issuerTradingSymbol>AMIX</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001662139</rptOwnerCik>
            <rptOwnerName>Hauser Bradley</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>C/O AUTONOMIX MEDICAL, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>21 WATERWAY AVENUE, SUITE 300</rptOwnerStreet2>
            <rptOwnerCity>THE WOODLANDS</rptOwnerCity>
            <rptOwnerState>TX</rptOwnerState>
            <rptOwnerZipCode>77380</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isOfficer>1</isOfficer>
            <officerTitle>CEO and President</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <derivativeTable>
        <derivativeTransaction>
            <securityTitle>
                <value>Option (Right to buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>1.35</value>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2024-06-17</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>900000</value>
                </transactionShares>
                <transactionPricePerShare>
                    <footnoteId id="F2"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F1"/>
            </exerciseDate>
            <expirationDate>
                <value>2034-06-17</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>900000</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>900000</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">The option set forth in this table vests in four equal annual installments beginning on the first anniversary of the Transaction Date, subject to the grantee's continued service to the Company on each vesting date.</footnote>
        <footnote id="F2">Issued in connection with the reporting person's employment with the Company.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Trent Smith, Attorney-in-Fact for Brad Hauser</signatureName>
        <signatureDate>2024-06-20</signatureDate>
    </ownerSignature>
</ownershipDocument>
